Method of detecting norwalk-like virus (gII)

ABSTRACT

A method of detecting a virus in a specimen, whereby a Norwalk-like virus (GII) is detected by using as an index the nucleic acids of a complementary nucleotide sequence corresponding to the 4851- to 5450-positions of the nucleotide sequence of the cDNA of the prototype (standard strain) of the Norwalk-like virus (GII); and a detection kit for performing this method.

TECHNICAL FIELD

[0001] The present invention relates to a method of detecting a virus.

BACKGROUND ART

[0002] The term “food poisoning” generally brings to mind bacterial foodpoisoning caused by bacteria such as Salmonella, Vibrioparahaemolyticus, and pathogenic E. coli, or natural toxin foodpoisoning caused by natural toxins contained in, for example, globefishor mushrooms. In addition, a very large number of food poisoning casesare caused by viruses, such as Norwalk-like viruses (hereinafterreferred to as NLVs), rotavirus, astrovirus, enterovirus, adenovirus,and hepatitis A virus. Recent epidemiological research has revealedthat, among other viruses, Norwalk-like viruses are typicalfood-poisoning viruses.

[0003] Norwalk virus was first identified in 1972 after an outbreak ofgastrointestinal illness in the U.S.A. Under an electron microscope, thevirus is observed as a small spherical virus of about 30 nm in diameterhaving an unclear surface structure, and since then viruses havingsimilar shapes have been collectively called “small round structuredviruses” (SRSVs). In the meantime, in 1974, calicivirus, which had beenwell known in veterinary medicine and which measures about 30 nm indiameter and assumes a unique surface structure resembling a “Star ofDavid,” was first identified in a human patient; specifically, in apatient suffering winter vomiting disease, which was at that timeepidemic in Britain. Since then, viruses having a shape similar to theabove have been called classical human caliciviruses.

[0004] These viruses are very difficult to grow in tissue culture cellsor in experimental animals, and therefore, for some time the onlyfeasible method was to isolate and culture the viruses on volunteers byuse of stool specimens. Thus, characterization of the viruses was quitedifficult. In 1990, a research group led by X. Jiang cloned the genomeof Norwalk virus, and since then, gene analysis of these viruses hasbeen energetically performed. Such efforts have revealed that an SRSVand a classical human calicivirus both belong to the family ofCaliciviridae, having a single stranded “plus” RNA (plus-stranded). Inthe XIth International Congress of Virology, the family Caliciviridaewas reported to comprise four different genera.

[0005] Thus, a group of viruses that had been called SRSVs wasdetermined to belong to the genus Norwalk-like viruses (NLVs), andanother group that had been called classical human caliciviruses wasdetermined to belong to the genus Sapporo-like viruses (SLVs) .Moreover, from an accumulation of data of genomic nucleotide sequencesof viruses collected from a vast number of clinical specimens, NLVs havebeen confirmed to be classified into two genogroups I and II; i.e.,genogroup I (GI) encompassing Norwalk viruses and Southampton viruses,and similar viruses; and genogroup II (GII) encompassing Hawaii viruses,Snow Mountain viruses, and similar viruses.

[0006] NLV infections in humans primarily occur by the mediation offoods (fish, shellfish, and water). Most of the viral food-poisoningcases that frequently occur during winter are believed to be caused byingestion of shellfish such as oysters, and in fact, in a great numberof study reports, oysters are identified as the source of infection withNLtVs. Some reports describe that ingestion of a sandwich contaminatedwith NLVs caused infection. Thus, presumably, NLV infection readilyspreads through feces from an infected patient. (This virus is known tohave strong infectivity and to cause infection even in a case whereseveral to about one hundred viruses are present in a food product).

[0007] Once food poisoning has occurred, needless to say, identificationof the cause and the contamination source is a critical issue. That is,the food-poisoning patients must be treated as quickly as possiblethrough appropriate selection of a therapeutic method, which would berealized by identifying the cause of the food poisoning, andsimultaneously, spreading of food poisoning must be stopped byidentifying the contamination source as early as possible.

[0008] In particular, in order to identify the cause and thecontamination source of food poisoning caused by pathogenicmicroorganisms, the following are required: detection and identificationof the pathogenic microorganism that caused the illness (i.e.,identification of the cause of the food poisoning); and identificationof the food and the food manufacturing facility that caused the foodpoisoning, on the basis of, for example, the diet history of the patientsuffering food poisoning (identification of the contamination source ofthe food poisoning).

[0009] Conventionally, an electron microscope has been employed todetect the above-mentioned NLVs. However, methods employing an electronmicroscope require an intricate procedure, and in addition, rapid andaccurate detection of viruses is difficult in cases where the quantityof the viruses is small. In particular, since a very small amount of NLVparticles exhibit infectivity, rapid and accurate detection of NLVs in,for example, contaminated foods is keenly desired, and yet, realizationhas been difficult Moreover, detection methods employing an electronmicroscope require a large facility for accommodating the electronmicroscope, and thus, detection through electron microscopy has beenpossible in only a limited number of facilities.

[0010] By keeping pace with the recent progress in gene analysistechniques, more sensitive, more rapid gene analysis through RT-PCR hasnow been performed frequently. In order to detect a virus through use ofthis method, primers for amplifying a specific region of the gene(hereinafter referred to as gene amplification primers) and primers usedin the process of detecting a gene amplification product of interest onthe basis of the presence of the specific region serving as an index(hereinafter referred to as detection primers) must be designed andemployed. Particularly in the case of viruses such as NLVs, design ofsuch primers encounters a problem which is very difficult to solve. Thatis, viruses easily undergo mutation and therefore, even in the case inwhich the virus responsible for the previous outbreak of food poisoningfalls within the same group of the virus that is now epidemic, there isa high risk that detection may be disabled unless primers different fromthose employed for detection of the virus in previous outbreaks are usedfor the current food poisoning. Needless to say, attaining accurateidentification of the source of contamination will still require use ofdetection primers each individually specific to viral mutation variants.However, this would only be required for the purpose of verification andwould suffice if performed after identification of the causative virusof the food poisoning and identification of the source of infection isalmost complete. More importantly, rapid identification should be givena high priority so as to establish a therapeutic regimen for thefood-poisoning patient and to prevent spreading of contamination.

[0011] In order to solve the above problem, a need exists fordiscovering a highly conserved region in genes of a virus of interest,and designing, among other things, detection primers which correspond tothe region, thus providing means for detecting the virus through use ofsuch tools.

[0012] Accordingly, an object of the present invention is to identify ahighly conserved region in genes of NLVS, and, on the basis of theinformation thus obtained, to provide rapid, accurate means fordetecting NLVs.

DISCLOSURE OF THE INVENTION

[0013] In order to attain the above object, the present inventors haveconducted careful studies and have successfully identified a highlyconserved region (bridging the vicinity of the C-terminus of the ORP 1region and the vicinity of the N-terminus of the ORF 2 region) in a geneof NLV genogroup II, and, on the basis of this finding, have devisedrapid, accurate means for detecting viruses belonging to genogroup II ofNLVS, among other NLVs, leading to completion of the present invention.

[0014] First, there will be described an essential discovery thatconstitutes the basis for the present invention; i.e., a highlyconserved region commonly found in genes of Norwalk-like viruses (NLVs)belonging to genogroup II (hereinafter also referred to as NLVs (GII)).

[0015] In order to identify the above-mentioned highly conserved regionin the gene of an NLV (GII), the present inventors performed thefollowing test.

[0016] Details of the Test

[0017] (1) Stool Specimens and Preparation of RNA Samples

[0018] Gene analysis of NLVs (GII) was performed on stool specimenscollected from 44 cases of non-bacterial gastroenteritis from which NLVparticles were detected through electron microscopy in the SaitamaInstitute of Public Health during 1998-2000.

[0019] Briefly, each of the stool specimens was suspended in sterilizeddistilled water so as to attain a concentration of about 10% (W/V), andthe suspension was subjected to centrifugation at 3000×g for 5 minutes.From the supernatant (140 μL), nucleic acid was extracted in accordancewith the manufacturer's protocol of an RNA extraction kit (QIA ViralRNA, Qiagen), and suspended in 50 μL sterilized distilled water, wherebyan RNA sample was obtained.

[0020] (2) Determination of Full Length Sequences of NLV Genes andAnalysis of the Genes

[0021] cDNA was synthesized from each of the thus-prepared RNA samplesby use of an oligo dT primer, and amplified through LongRT-PCR. Thenucleotide sequence of each of the gene amplification products wasdetermined through direct sequencing by use of the primer walking method(Nucleic Acids Res. 1989 17(15): 6087-6102). Sequencing of the genomic5′-terminus was performed using three types of RACE (rapid amplificationof cDNA ends).

[0022] Through the gene analysis employing the above RNA samples, theentire nucleotide sequences of 8 new NLV strains (#U1/AB039775,#U3/AB039776, #U4/AB039777, #U16/AB039778, #U17/AB039779, #U18/AB039781,#U25/AB039780, #U201/AB039782) were determined, and partial nucleotidesequences (53 sequences) of 33 strains were also determined (#17, #19a,#19b, #19c, #19d, #19f, #19e, #82, #83, #84a, #84b, #89, #105a, #105b,#105c, #105d, #105e, #16, #101, #111a, #111b, #18, #26, #27a, #27b, #31,#32, #34, #35, #36, #37a, #37b, #37c, #53a, #53b, #62, #63, #66, #68,#80a, #80b, #80c, #80d, #80e, #80f, #109, #85, #88, #9, #49, #93, #98,#99). Genome diversity was investigated by use of the nucleotidesequences of a number of NLV variants, including these new variants, aprototype (standard strain; AF145896/Camberwell), and known NLV (GII)variants which had already been registered in Genbank (X86557/Lorsdale,X81879-2/melksham, U07611-2/Hawaii, U02030-2/Tronto(TV24),L23830-2/OTH-25/89/J, AF190817-2/Arg320, X76716-1/Bristol, U22498-2/MK,U46039 Auckland, U75682-1/Snow Mountain Strain, AB032758 Chitta virus,AF080549-1/1996/SC, AF080550-1/345-2/96002737/1996/,AF080551-1/004/95M-14/1995/AU,AF080552-1/358/96015107/1996/FL,AF080553-1/364/96019537/1996/AZ, AF080554-1/366/96019554/1996/ID,AF080555-1/373/96019743/1996/SC, AF080556-1/379/96019984/1996/AZ,AF080557-1/384/96025046/1996/FL, AF080558-1/408/97003012/1996/FL,AF080559-1/416/97003156/1996/LA, AF195847-1/Alphatron/98-2/1998/,AF195848-1/Amsterdam/98-18/1998, AJ004864-1/Grimsby,HCA277606/Girlington/93/UK,HCA277607/Hillingdon/90/UK,HCA277608/Leeds/90/UK, HCA277611/Bhaml32/95/UK,HCA277613/Parkroyal/95/UK, HCA277617/Rbh/93/UK, HCA277618/Wortley/90/UK,HCA277619/Symgreen/95/UK, HCA277620/Seacroft/90/UK, 070059-1/SnowMountain strain, AB005260-1/SA1/89/Japan, AB005261-1/SA2/91/Japan,AB020547-1/TOB-93-Japan, AB020549-1/TOC1-93-J,AB020551-1/TOC2-93-J,AB020552-1/IZ10-94-J,AB020563-1/MH22-82-J,AB028244/NLV36, AB028245/NLV21, AB028246/NLV114), and the most highlyconserved gene region was searched.

[0023] In the present invention, the reference employed as a basis fordescribing a gene region is the nucleotide sequence (cDNA sequence) of agene of the above-mentioned prototype (standard strain),AF145896/Camberwell. FIG. 1 shows the results of the investigation ofgenome diversity. In chart i) of FIG. 1, the X-axis represents the basenumber—as counted from the 5′-terminus—of the gene (cDNA) of theabove-mentioned prototype of NVLs (GII), and the Y-axis represents thedegree of conservation (the greater the Y-axis value, the more analogousthe nucleotide sequences of respective strains, meaning that the gene ishighly conserved, and conversely, the smaller the Y-axis value, the morevaried the nucleotide sequences of respective strains, meaning that thegene is less conserved). Chart ii) of FIG. 1 shows functions, in NLVs(GII), of the gene having the above-described nucleotide sequences.

[0024] Analysis of the genes of NLVs (GII) shown in FIG. 1 revealed thatthe region in which the genes from respective strains exhibit thehighest homology lies between the vicinity of the C-terminus of the ORF1region and the vicinity of the N-terminus of the ORF2 region, where themaximum value of homology was found to be 90% or more.

[0025] FIGS. 2 to 5 show, in an orderly arranged form for the purpose ofcomparison, nucleotide sequences of the respective strains of NLVs(GII), spanning from the vicinity of the C-terminus of ORF1 (FIGS. 2A to2C) to the vicinity of the N-terminus of ORF2 (FIGS. 4A and 4D, and 5Ato 5C; Note that FIGS. 3A to 3C show nucleotide sequences of a regionthat bridges ORF1 and ORF2). In FIGS. 2 to 5, the names of therespective strains of NLVs (GII) employed are shown in the left column,and the nucleotide sequence of the prototype (standard strain) is shownin the uppermost row, As described hereinabove, in the presentinvention, the base number of the prototype shown in the uppermost rowis employed as a reference. In the nucleotide sequences of respectivestrains (excepting the prototype), the symbol “.” represents that thebase at that position is the same as that of the prototype (standardstrain), and the vacancies indicate that the bases found at thecorresponding positions in the prototype are absent. The symbol “*” inthe lowermost row indicates that the base at that position is in commonthroughout the strains, and the symbol “.” in the lowermost rowindicates the presence of any difference in base among the strains.

[0026] (3) Conclusion

[0027] The investigation on gene conservation of NLVs (GII) hasclarified that the nucleotide sequence region exhibiting geneconservation of such a degree that enables use of that region indetection of NLVs (GII) is a region corresponding to the 4851- to5450-positions of the nucleotide sequence of the cDNA of the prototype(standard strain) of NLVs (GII); that a region corresponding to the4919- to 5389-positions of the nucleotide sequence exhibits aparticularly high level of conservation, and a region corresponding tothe 4988- to 5107-positions of the nucleotide sequence exhibits anexceptionally high conservation (in particular, a region correspondingto the 5042- to 5067-positions exhibits the highest conservation(hereinafter this region may be referred to as the significantly highlyconserved region). (Within this context, the expression “correspondingto” is used to describe a relation between two corresponding nucleotidesequence regions, one being from the cDNA of the above-mentionedprototype and the other being from a variant, which relation iselucidated through gene analysis of NLVs (GII), including variants.Specifically, the expression “corresponding to” is used to refer to theincidence in which the nucleotide sequence in question can hybridizewith nucleic acids of the aforementioned nucleotide sequence of the cDNAof the prototype under stringent conditions (56 to 68° C., in thepresence of 50 mM or more sodium ions). Specific examples includenucleotide sequences of NLVs (GII) shown in FIGS. 2 to 5, whichcorrespond to the gene region represented by the above-mentionednucleotide sequence of the cDNA of the prototype.]

[0028] The present invention is directed to means for rapidly andaccurately detecting NLVs (GII) by making use of the nucleotide sequenceof a nucleic acid in a highly conserved gene region (i.e., the regioncorresponding to the 4851- to 5450-positions of the nucleotide sequenceof the cDNA of the prototype (standard strain) of NLVs (GII),hereinafter such a gene region is also referred to as a “conservedregion”), which is determined through gene analysis regarding geneconservation of NLVs (GII). Specifically, the present invention providesa viral detection method for Norwalk-like viruses (GII) in a specimen(hereinafter also referred to as “the present detection method”) by useof, as an index, the nucleic acid fragment of a complementary nucleotidesequence or complementary nucleotide sequences (hereinafter collectivelycalled “a complementary nucleotide sequence”) corresponding to the 4851-to 5450-positions (preferably 4919- to 5389-positions (hereinafter alsoreferred to as the highly conserved region), more preferably 4988- to5107-positions (hereinafter also referred to as the special conservedregion)) of the nucleotide sequence of the cDNA of the prototype(standard strain) of NLVs (GII).

[0029] As used herein, the word “complementary” is used to describe arelation where a nucleic acid fragment having a certain nucleotidesequence exhibits such a degree of complementation that enableshybridization with another nucleic acid fragment under stringentconditions (56 to 68° C., in the presence of 50 mM or more sodium ions),and the expression “complementary nucleotide sequence” encompasses thefollowing incidences: The “complementary nucleotide sequence” isminus-stranded with respect to a plus-stranded fragment of thenucleotide sequence; the “complementary nucleotide sequence” isplus-stranded with respect to a minus-stranded fragment of thenucleotide sequence; and one “complementary nucleotide sequence” isminus-stranded with respect to a plus-stranded fragment of thenucleotide sequence and the other “complementary nucleotide sequence” isplus-stranded with respect to a minus-stranded fragment of thenucleotide sequence.

BRIEF DESCRIPTION OF THE DRAWINGS

[0030]FIG. 1 shows the results of the investigation of genome diversityof NLVs (GII).

[0031]FIGS. 2A to 2C show nucleotide sequences identified in respectivestrains of NLVs (GII), arranged for facilitating comparison, thesequences coding for the vicinity of the C-terminus of the ORF1 region.

[0032]FIGS. 3A to 3C show nucleotide sequences identified in respectivestrains of NLVs (GII), arranged for facilitating comparison, thesequences coding for the region from the vicinity of the C-terminus ofthe ORF1 region to the vicinity of the N-terminus of the ORF2 region.

[0033]FIGS. 4A to 4C show nucleotide sequences identified in respectivestrains of NLVS (GII), arranged for facilitating comparison, thesequences coding for the vicinity of the N-terminus of the ORF2 region(part 1 of 2).

[0034]FIGS. 5A to 5C shows nucleotide sequences identified in respectivestrains of NLVs (GII), arranged for facilitating comparison, thesequences coding for the vicinity of the N-terminus of the ORF2 region(part 2 of 2).

BEST MODE FOR CARRYING OUT THE INVENTION

[0035] Modes for carrying out the present invention will next bedescribed.

[0036] In order to make use of the finding on conserved regions of NLVs(GII) as an index for detecting NLVs (GII), any of the mentionedconserved regions (hereinafter collectively referred to as “ConservedRegions”; including the “highly conserved region,” “special conservedregion,” and “significantly highly conserved region”; the conventionapplies hereafter) must be amplified through a nucleic acidamplification method, to thereby produce gene amplification products ofthe Conserved Regions. Examples of such an amplification method includePCR and other methods, such as RT-PCR, NASBA (nucleic acid sequencebased amplification), and SDA (strand displacement amplification). Inany of these methods, gene amplification primers must be designed andprepared so as to be adapted to the method. Generally, geneamplification primers are designed such that a gene region is amplifiedin both directions of forward (5′→3′) and reverse, with the gene regionbeing sandwiched by +/−primers. Since the gene amplification primersmust properly bind to the intended regions of the gene serving as atemplate for attaining complementary binding, it is necessary that anyof such primers contain at least a minimum number of bases forming anucleotide sequence complementary only to characteristic nucleotidesequence(s). Such a gene amplification primer has at least 10 bases,preferably about 15 to 30 bases. In addition, for 5 bases counted fromthe 3′-terminus in a region that is to achieve a complementary binding,the primer is preferably designed to be as precisely complementary aspossible.

[0037] Moreover, the gene region which is employed as a basis forestablishing gene amplification primers must at least meet therequirement that the resultant gene amplification product contains acomplementary nucleotide sequence corresponding to Conserved Regions.From this viewpoint, a preferred gene amplification primer comprises acomplementary nucleotide sequence corresponding to consecutive 10 ormore bases, preferably consecutive 15 to 30 bases, of a nucleotidesequence selected, as a benchmark, from the group consisting ofnucleotide sequences of the 4988- to 5028-positions of the nucleotidesequence of the cDNA of the prototype (standard strain) of NLVs (GII),the 5080- to 5107-positions thereof, the 4919- to 4941-positionsthereof, and the 5367- to 5389-positions thereof.

[0038] Nucleic acids having nucleotide sequences that correspond to aconserved region—the nucleic acids serving as indices in detection ofNLVs (GII) in the present invention—are obtained as follows: at leasttwo nucleotide sequences having 10 or more consecutive bases (preferablya complementary nucleotide sequence having 15 to 30 bases) are selectedfrom the conserved region; and through use of gene amplification primersestablished on the basis of the selected nucleotide sequences, genesobtained from specimens are subjected to the above-mentionedamplification means, to thereby yield the target nucleic acids as thegene amplification products. Preferably, the gene amplification productsinclude a region corresponding to the 4942- to 5366-positions of thenucleotide sequence of the cDNA of the prototype of NLVs (GII), which isa highly conserved region within the aforementioned conserved regions.Most preferably, the nucleotide sequence to be included in the geneamplification products obtained from the highly conserved region is asegment corresponding to the 5042- to 5067-positions of thecomplementary nucleotide sequence of the cDNA of the prototype of NLVs(GII), which is the significantly highly conserved region.

[0039] That is, through use of the gene amplification primers of any ofthe above combinations, the resultant gene amplification productsoriginating from NLVs (GII) will contain at least a nucleotide sequencecorresponding to the 5042- to 5067-positions of the nucleotide sequenceof the cDNA of the prototype of NLVs (GII). Therefore, through use ofMeans which enables detection of the nucleotide sequences of thementioned highly conserved regions, most typically, through use ofnucleic acid probes complementary to a complementary nucleotide sequencecontaining a portion of or the entirety of the highly conserved regionsfalling within the nucleic acid of the detection target, detection andquantitation of the resultant gene amplification product, which is thetarget of detection, can be attained, and using thequantitation/detection results as indices, NLVs (GII) can be detectedrapidly and accurately.

[0040] Notably, in order to obtain a gene amplification productcontaining a nucleotide sequence of the significantly highly conservedregion, the at least two nucleotide sequences based on which geneamplification primers are established, each nucleotide sequence beingcomposed of 10 or more consecutive bases, are preferably selected fromthe cDNA nucleotide sequence set of 4988- to 5028-positions and 5080- to5107-positions of the nucleotide sequence of the cDNA of the prototype(standard strain) of Norwalk-like viruses (GII).

[0041] Through use of the gene amplification primers falling within theabove combinations, the resultant gene amplification productsoriginating from NLVS (GII) will contain a nucleotide sequencecorresponding to the 5042- to 5067-positions of the nucleotide sequenceof the cDNA of the prototype of NLVs (GII). Therefore, through use ofmeans which enables detection of the nucleotide sequences of thementioned significantly highly conserved regions, most typically,through use of nucleic acid probes complementary to a complementarynucleotide sequence containing a portion of or the entirety of thesignificantly highly conserved regions falling within the nucleic acidof the detection target, detection and quantitation of the resultantgene amplification product, which is the target of detection, can beattained, and using the quantitation/detection results as indices, NLVs(GII) can be detected rapidly and accurately.

[0042] Each of the above-described gene amplification primers may beemployed as a component of the detection kit of the present inventiondescribed hereinbelow.

[0043] Notably, the present detection method can be applied to anyspecimen in which NLVS (GII) are to be detected. For example, when NLVs(GII) are to be detected in a food-poisoning patient, stool of thepatient is typically employed as a specimen, and depending on the case,vomitus, blood, etc. may be employed. When NLVs (GII) are to be detectedin food or a food production facility, the food itself, depositscollected from the food production facility, clothing of food productionworkers, etc. may be employed as a specimen. Moreover, the specimen maybe obtained from watery sources, such as various types of sewage ordischarging water, seawater, river water, and lake water. In order toobtain genes from the aforementioned specimens, suitable methods areselected according to the type of the specimen to be employed.Generally, a specimen is immersed or suspended in water or a similarmedium, and from a supernatant fraction obtained therefrom, viral RNA isextracted, through a conventional method such as the acid phenol method(e.g., the acid guanidinum-phenol-chloroform (AGPC) method). Thethus-obtained viral RNA is processed through a suitably selected geneamplification method. For example, through preparation of cDNA having anucleotide sequence complementary to the nucleotide sequence of theviral RNA using a reverse transcriptase—to thereby obtain a nucleic acidsample. The nucleic acid sample is then subjected to gene amplificationby use of the aforementioned gene amplification primers, whereby a geneamplification product of interest can be obtained (generally, thepresence of the product of interest can be confirmed throughelectrophoresis on the basis of size of the target product).

[0044] Conventional means may be employed to detect, as an index of thepresence of NLVs (GII), a certain specific nucleotide sequence of a geneamplification product. Typically, through use of nucleic acid probes fordetection (for example, nucleic acid fragments which have been labeledwith a fluorophore or a radioisotope) containing nucleotide sequencescomplementary to the aforementioned complementary nucleotide sequence(s)that serve(s) as an index of virus detection, the presence or absence ofthe nucleotide sequence of the gene amplification product can beconfirmed by hybridization or a similar method (as a negative result orpositive result). Generally, such detection means is performed on theamplification product after the gene amplification process hascompleted. However, such a procedure involves a drawback in that since atechnician must open a tube to remove the analytical sample after geneamplification reaction, there will be increased chances whereexperimental facilities or reagents may be polluted with the geneamplification product, and in addition, extra time and labor may berequired. Moreover, the polluting gene amplification product may invitethe risk of false positive results, raising critical problems in thefield of detection. Accordingly, in order to avoid risks such aspollution and to minimize the time required for detection, it isrecommended to use, during gene amplification, means enabling monitoringof the presence of a specific nucleotide sequence of the geneamplification product. Examples of typical methods employing such meansinclude, but are not limited to, 1) a detection method by use of amolecular beacon probe, and 2) a detection method by use of a Taq-Manprobe.

[0045] 1) The detection method by use of a molecular beacon probe makesuse of a hair-pin shaped hybridization probe (molecular beacon probe)for allowing fluorescent monitoring of a gene amplification productobtained by PCR during or after an amplification procedure (NatureBiotechnology, 1998, 16: 49-53). Terminal sequences of the nucleic acidfragment constituting the molecular beacon probe are complementary toeach other. Typically, the terminal portions are bonded to each other,whereby a “stem-loop structure” is formed. The loop portion of thestem-loop structure is designed so as to be complementary to a region ofinterest (i. ., Conserved Regions) of the gene amplification product.Moreover, to one end of the nucleic acid of the probe, a fluorophore isbound, and to the other end of the nucleic acid of the probe, anon-fluorescent quencher dye is bound. When the probe is present in asolution in a free state, the probe has a hairpin structure, whereby thefluorophore and the quencher interact each other, and fluorescence isnot detected. However, when the solution contains the gene amplificationproduct having a nucleotide sequence complementary to the nucleotidesequence of the probe, the loop portion is bound to the complementarynucleotide sequence portion. As a result, the overall structure of theprobe varies, resulting in a separation of the fluorophore and thequencher from each other, and thus quenching effect of the quencher forthe fluorophore is canceled. Therefore, fluorescence emitted from thefluorophore can be observed. Increase in fluorescence intensity causedby canceling of the quenching effect is proportional to the increment inthe amount of the gene amplification product having the nucleotidesequence complementary to the sequence of the nucleic acid that formsthe probe. Through monitoring the increase in the fluorescenceintensity, the presence of the target nucleotide sequences (e.g.,sequences of Conserved Regions) can be detected not only aftercompletion of gene amplification, but also during gene amplification.That is, NLVs (GII) in a specimen can be detected through use of theresults of the aforementioned increase in fluorescence intensity as anindex.

[0046] Labeling of a molecular beacon probe through use of theaforementioned fluorophore and the quencher is typically performed asfollows. The 5′-terminus of the nucleic acid probe is labeled with afluorescein fluorophore (such as 6-carboxyfluorescein (6-FAM) or6-carboxy-4,7,2′, 7′-tetrachlorofluorescein (TET)) or a rhodaminefluorophore (such as 5-carboxytetramethylrhodamine (TAMARA)), and the3′-terminus of the probe is labeled with a quencher (such as4-(4′-dimethylaminophenylazo)benzoic acid (DABCYL)) (see, for example,Nature Biotechnology, 1996, 14: 303-308).

[0047] 2) The detection method by use of a Taq-Man probe makes use of ahybridization probe (Taq-Man probe) for allowing fluorescent monitoringof a gene amplification product obtained by PCR during the amplificationprocedure (see, for example, Experimental Medicine, Vol. 15, No. 7(extra issue), pp. 46-51, 1997). The Taq-Man probe is a nucleic acidfragment bearing a fluorescein fluorophore label (reporter dye) and arhodamine fluorophore label (quencher dye) at the 5′-terminus and3′-terminus of the fragment, respectively. When the reporter dye and thequencher dye are linked to each other via the nucleic acid fragment, dueto Forster resonance energy, the quencher dye inhibits fluorescenceemission of the reporter dye. However, as elongation proceeds along withthe progress of annealing of a nucleic acid (in the gene amplificationproduct) which is complementary to the nucleic acid of the Taq-Man probeperformed through use of primers and the Taq-Man probe, hydrolysisoccurs from the 5′-terminus of the Taq-Man probe under 5′→3′endonuclease activity of Taq DNA polymerase. As a result, the5′-terminal reporter dye is released from the 3′-terminal quencher dye,resulting in an increase in fluorescence intensity of the reporter dyewhich has been suppressed. The increase in fluorescence intensity causedby the reporter dye is proportional to the increase in the amount of thegene amplification product having the nucleotide sequence complementaryto the sequence of the nucleic acid that forms the probe Throughmonitoring the increase in fluorescence intensity, the presence of thenucleotide sequence of interest (e.g., Conserved Regions) can bedetected not only after completion of gene amplification, but alsoduring gene amplification. Thus, NLVs (GII) in a detection specimen canbe detected through use of the results of the aforementioned increase influorescence intensity as an index.

[0048] Labeling of a Taq-Man probe with the aforementioned fluorophoreis typically performed in accordance with a conventional method bylabeling the 5′-terminus of the probe with a fluorescein fluorophore(such as 6-FAM or TET), and the 3′-terminus of the probe with arhodamine fluorophore (such as TAMARA) (see, for example, Nucleic AcidsResearch, 1993, 21 (16): 3761-3776).

[0049] All the aforementioned nucleic acid probes to be used fordetection purposes (hereinafter also referred to as nucleic acid probesfor detection) may be employed as components of the detection kit of thepresent invention which will be described below.

[0050] In relation to the detection of NLVs (GII) according to thepresent detection method, detection of a gene amplification product or asimilar product having a nucleotide sequence of interest may beperformed by way of quantitation of the nucleic acid fragment ofinterest through use of the aforementioned means. Alternatively, insteadof quantitation, NLVs (GII) may be detected in the form of qualitative(e.g., positive or negative) information. By use of the detectionresults of the gene amplification products (quantitative value orqualitative information) as an index, and by correlating the index tothe presence/absence or the amounts of the NLVs (GII) in the specimen,the NLVs (GII) of interest can be detected.

[0051] The present invention also provides a kit for detecting NLVs(GII) (hereinafter, the kit will be referred to as the present detectionkit) for performing the present detection method.

[0052] The present detection kit typically contains primers foramplifying—by RT-PCR or any other suitable method—a nucleic acidfragment having any of Conserved Regions of NLVs (GII), and/or one ormore probes for detecting the Conserved Regions of a gene amplificationproduct. Preferably, the kit contains both; primers and probes.

[0053] The gene amplification primers are nucleic acid fragments capableof producing gene amplification products through amplification of aConserved Region of an NLV (GII) gene by use of gene amplification meanssuch as RT-PCR. The nucleic acid probes for detection contain nucleicacid fragments having nucleotide sequences complementary to sequencescorresponding to the nucleotide sequences of a Conserved Region. Asdescribed above, one may employ nucleic acid probes for detection of abasic mode comprising complementary nucleotide sequences labeled with afluorophore or a radioisotope. In particular, when NLVs (GII) aredetected in the course of gene amplification, preferably, there may beemployed such nucleic acid probes for detection produced byincorporating nucleic acids of a complementary nucleotide sequence intothe aforementioned molecular beacon probe or Taq-Man probe.

[0054] Details of the gene amplification primers and the nucleic acidprobes for detection which may be included in the present detection kithave already been described in relation to the present detection method.Specific examples of such primers and probes are also described in theExample section below.

EXAMPLE

[0055] The present invention will next be described by way of example,which should not be construed as limiting the technical scope of theinvention.

[0056] [Preparation of Primers and Probes]

[0057] Detection of NLVs (GII) in the present example was performed byuse of the following nucleic acid primers for gene amplification andnucleic acid probes for gene detection. The nucleic acids describedbelow were chemically synthesized by use of a full-automated ABI 3948Nucleic Acid Synthesis and Purification System (Applied Biosystems).

[0058] Gene Amplification Primers

[0059] [Forward Primers (Used in the Form of a Mixture or Individually]

[0060] For amplification of a special conserved region:

[0061] G2FA: 5′CARGARBCNATGTTYAGRTGGATGAG 3′ (SEQ ID NO; 1,corresponding to the 5003- to 5028-positions of the nucleotide sequenceof the prototype)

[0062] For amplification of a highly conserved region:

[0063] G2FB1: 5′GGHCCMBMDTTYTACAGCAA 3′ (SEQ ID NO: 2, corresponding tothe 4922- to 4941-positions of the nucleotide sequence of the prototype)

[0064] G2FB2: 5′GGHCCMBMDTTYTACAAGAA 3′ (SEQ ID NO: 3, corresponding tothe 4922- to 4941-positions of the nucleotide sequence of the prototype)

[0065] G2FB3: 5′GGHRCCMBMDTTYTACARNAA 3′ (SEQ ID NO: 4, corresponding tothe 4922- to 4941-positions of the nucleotide sequence of the prototype)

[0066] [Reverse Primers (Used in the Form of a Mixture or Individually]

[0067] For amplification of a special conserved region:

[0068] G2RA1: 5′TCGACGCCATCTTCATTCACA 3′ (SEQ ID NO: 5, corresponding toa nucleotide sequence that is complementary to the 5080- to5100-positions of the nucleotide sequence of the prototype)

[0069] G2RA2: 5′TCAYTCGACGCCATCTTCAT 3′ (SEQ ID NO: 6, corresponding toa nucleotide sequence that is complementary to the 5085- to5104-positions of the nucleotide sequence of the prototype)

[0070] For amplification of a highly conserved region:

[0071] G2RB1: 5′CCACCWGCATAACCATTRTACAT 3′ (SEQ ID NO: 7, correspondingto a nucleotide sequence that is complementary to the 5367- to5389-positions of the nucleotide sequence of the prototype)

[0072] G2RB2: 5′CCACCWGCATGCCCATTRTACAT 3′ (SEQ ID NO: 8, correspondingto a nucleotide sequence that is complementary to the 5367- to5389-positions of the nucleotide sequence of the prototype)

[0073] G2RB3: 5′CCRCCNGCATRHCCRTTRTACAT 3′ (SEQ ID NO: 9, correspondingto a nucleotide sequence that is complementary to the 5367- to5389-positions of the nucleotide sequence of the prototype)

[0074] Nucleic Acid Probes for Gene Detection

[0075] [Taq-Man Probes]

[0076] G2TM1: 5′TGGGAGGGCGATCGCAATCT 3′ (SEQ ID NO: 10, corresponding tothe 5048- to 5067-positions of the nucleotide sequence of the prototype)

[0077] G2TM2: 5′AGATTGCGATCGCCCTCCCA 3′ (SEQ ID NO: 11, corresponding toa nucleotide sequence that is complementary to the 5048- to5067-positions of the nucleotide sequence of the prototype)

[0078] The 5′-terminus and the 3′-terminus of all the Taq-Man probesused in the present example were labeled with TET and TAMARA,respectively (labeling was performed in accordance with the methoddescribed in Nucleic Acids Research (1993, 21 (16): 3761-3766)).

[0079] [Molecular Beacon Probes (the Lowercase Letters Denote StemPortions)]

[0080] G2MB1: 5′ccgtCgTGGGAGGGCGATCGCAATCTcgacgg 3′ (SEQ ID NO: 10,corresponding to the 5048- to 5067-positions of the nucleotide sequenceof the prototype)

[0081] G2MB2: 5′cgtgctGGGAGGGCGATCGCAAgcacg 3′ (SEQ ID NO: 12,corresponding to the 5049- to 5064-positions of the nucleotide sequenceof the prototype)

[0082] G2MB3: 5′ccgtcgAGATTGCGATCGCCCTCCCAcgacgg 3′ (SEQ ID NO: 11,corresponding to a nucleotide sequence that is complementary to the5048- to 5067-positions of the nucleotide sequence of the prototype)

[0083] G2MB4: 5′ccgtcgATTGCGATCGCCCTCCCAcgacgg 3′ (SEQ ID NO: 12,corresponding to a nucleotide sequence that is complementary to the5050- to 5067-positions of the nucleotide sequence of the prototype)

[0084] G2MB5: 5′ccgtggATTGCGATCGCCCTCCCccacgg 3′ (SEQ ID NO: 13,corresponding to a nucleotide sequence that is complementary to the5048- to 5066-positions of the nucleotide sequence of the prototype)

[0085] G2MB6: 5′cgtggaATTGCGATCGCCCTCCCtccacg 31 (SEQ 10 NO: 13,corresponding to a nucleotide sequence that is complementary to the5048- to 5066-positions of the nucleotide sequence of the prototype)

[0086] G2MB7: 5′cctgcATTGCGATCGCCCTCCCAgcagg 3′ (SEQ ID NO: 12,corresponding to a nucleotide sequence that is complementary to the5048- to 5067-positions of the nucleotide sequence of the prototype)

[0087] The 5′-terminus and the 3′-terminus of all the molecular beaconprobes used in the present example were labeled with TET and DABCYL,respectively (labeling was performed in accordance with the methoddescribed in Nature Biotechnology (1996, 14: 303-308)).

[0088] [Detection of virus]

[0089] (1) The detection method of the present invention was performedon the RNA samples extracted from stool specimens collected from 44cases of non-bacterial gastroenteritis from which NLV particles weredetected through electron microscopy performed in the Saitama Instituteof Public Health during the period from 1998 to 2000, the specimensbeing the same as those employed in the above-described test toinvestigate highly conserved regions of NLv (GII) genes.

[0090] First, each of the RNA samples was subjected to reversetranscription reaction. Briefly, each of the RNA samples (8 μL) wasmixed with a 12 μL solution for reverse transcription reaction. (Thesolution was prepared as follows: A dNTP solution (10 mM, 1 μL), randomhexamer (75 pmol), RNasin (30 units, Promega, USA), SuperScript IIRNaseH (−) Reverse Transcriptase (200 units, Gibco BRL, USA), DTT (100mM, 1 μL), and a 5-fold diluted reverse transcription buffer (250 mM,Tris-HCl (pH 8.3), 375 mM KCl, 15 MM MgCl₂) were mixed, and the mixturewas diluted with sterilized distilled water to attain a total volume of12 μL.) The mixture was allowed to react at 42° C. for one hour or more.Subsequently, the resultant mixture was subjected to enzyme inactivationreaction for 15 minutes at 70° C., whereby a cDNA sample correspondingto each of the RNA samples (RT products) was prepared. Separately, areaction mixture was prepared as follows: A buffer (25 μL), RT products(5 μL), primer (50 nM each), and fluorescent probe (Taq-Man probe G2TM1,5-20 pmol) were mixed, and the mixture was diluted with sterilizeddistilled water to attain a total volume of 50 μL. The reaction mixturewas subjected to PCR reaction by use of a nucleic acid obtained throughmixing G2FA, G2RA1, and G2RA2 (gene amplification primers) as a primerset for gene amplification, and a Taq-Man universal buffer Kit (ABI,USA) (PCR cycle: 50° C. for two minutes→95° C. for 10 minutes→(95°Csec→56° C. for three minutes)×50 cycles)). The fluorescence intensitywas monitored over time during the reaction by use of ABI7700 (ABI,USA).

[0091] A cDNA fragment of SzuGII strain was subjected to cloning by useof pT7blue vector (Novagen, USA) through a conventional method. Thefragment has a nucleotide sequence corresponding to the 5207- to5696-positions of the nucleotide sequence of the cDNA of the prototype(standard strain) of NLVs (GII). Through use of the thus-cloned cDNA asa positive control specific to NLVs (GII), NLV (GII) gene detection wasperformed in accordance with the aforementioned procedure. In all thecases in which the primers and probes of Set A or B were used, thefollowing were confirmed: 10¹ to 10⁷ copy genes of NLVs (GII) can bedetected (detection limit: 10¹ copy genes/reaction), and NLVs (GII) canbe quantitatively detected by reference to Ct values (threshold cyclenumber).

[0092] Among the aforementioned stool specimens which had been collectedfrom 44 patients of non-bacterial gastroenteritis, 28 cases (63.6%) werefound to be NLV (GII) positive, whereas the remaining 16 cases (36.4%)were found to be NLV (GII) negative. However, in all these 16 cases,WLVs (GI) were detected by another method. Taking the results regardingthese two types of NLVs (GII and GI) together, the detection rate ofNLVs was found to be 100%.

[0093] In cases where a system similar to the above and the followingcombination of gene amplification primers were used, results similar tothose described above were obtained: (combination: a mixture of G2FB1,G2FB2, G2FB3, G2RA1, and G2RA2; and a mixture of G2FA, G2RB1, G2RB2, andG2RB3).

[0094] Also, when the gene detection probe was a Taq-Man probe G2TM2(instead of G2TM1), or a molecular beacon probe G2MB1 to 7, analogousresults were obtained.

[0095] (2) In order to demonstrate that the detection method of thepresent invention can be carried out on food, the method was performedthrough use of fresh oysters instead of the aforementioned stoolspecimens as test specimens.

[0096] From each of the 40 fresh oyster individuals, the midgut glandwas removed, and sterilized distilled water was added thereto.Subsequently, the midgut gland was subjected to three cycles of freezingand thawing, whereby the tissues of the midgut gland were lyzed. Thelyzed tissues were subjected to centrifugation at 10,000×g for 20minutes. From the supernatant (140 μL) obtained through thecentrifugation, nucleic acid was extracted through use of QIA Viral RNA(QIAGEN, USA) in accordance with the manufacturer's protocol. Theextract was suspended in sterilized distilled water (50 μL). Through useof the suspension as an RNA sample of each fresh oyster, the detectionmethod of the present invention was carried out by use of a molecularbeacon probe G2MB1 as a gene detection probe in accordance with theprocedure described in (1) above.

[0097] As a result, NLVs (GII) were detected in two fresh oysterindividuals.

[0098] The results obtained in (1) and (2) clarified that the detectionmethod of the present invention enables rapid, accurate detection ofNLVs (GII) Moreover, as described above, through use of a nucleic acidprobe specific to NLVs (GII) as a primer for gene detection, bymonitoring the gene amplification process, the detection method of thepresent invention enables quantitative detection of NLVs (GII). Inconclusion, the present invention provides a virus detection method ofvery high sensitivity and efficiency.

Industrial Applicability

[0099] The present invention has identified a highly conserved generegion in the genes of NLVs (GII), and on the basis of this finding, arapid, accurate means for detecting NLVs is provided.

1 125 1 26 DNA Norwalk-like Virus (GII) misc_feature (9)..(9) n is a, c,g, or t 1 cargarbcna tgttyagrtg gatgag 26 2 20 DNA Norwalk-like Virus(GII) 2 gghccmbmdt tytacagcaa 20 3 20 DNA Norwalk-like Virus (GII) 3gghccmbmdt tytacaagaa 20 4 20 DNA Norwalk-like Virus (GII) misc_feature(18)..(18) n is a, c, g, or t 4 gghccmbmdt tytacarnaa 20 5 21 DNANorwalk-like Virus (GII) 5 tcgacgccat cttcattcac a 21 6 20 DNANorwalk-like Virus (GII) 6 tcaytcgacg ccatcttcat 20 7 23 DNANorwalk-like Virus (GII) 7 ccaccwgcat aaccattrta cat 23 8 23 DNANorwalk-like Virus (GII) 8 ccaccwgcat gcccattrta cat 23 9 23 DNANorwalk-like Virus (GII) misc_feature (6)..(6) n is a, c, g, or t 9ccrccngcat rhccrttrta cat 23 10 20 DNA Norwalk-like Virus (GII) 10tgggagggcg atcgcaatct 20 11 20 DNA Norwalk-like Virus (GII) 11agattgcgat cgccctccca 20 12 27 DNA Artificial Sequence probe 12cgtgctggga gggcgatcgc aagcacg 27 13 29 DNA Artificial Sequence probe 13ccgtggattg cgatcgccct cccccacgg 29 14 600 DNA Norwalk-like Virus (GII)14 ctgaaacaat gattccacac tcccaaagac ccatacaatt aatgtcccta ctgggagagg 60ccgcactcca cggcccagca ttctacagca aaattagcaa gctagtcatt gcagagttga 120aggaaggtgg catggacttt tacgtgccca gacaagagcc aatgttcaga tggatgagat 180tctcagatct gagcacgtgg gagggcgatc gcaatctggc tcccagtttt gtgaatgaag 240atggcgtcga gtgacgccaa cccatctgat gggtccgcag ccaacctcgt cccagaggtc 300aacaatgagg ttatggctct ggagcccgtt gttggtgccg ctattgcggc acctgtagcg 360ggccaacaaa atataattga cccctggatt agaaataatt ttgtacaagc ccctggtgga 420gagtttacag tgtcccctag aaacgctcca ggtgagatac tatggagcgc gcccttgggc 480cctgatttga atccctatct ttctcacttg tccagaatgt ataatggtta tgcaggtggt 540tttgaagtgc aagtgatcct tgcggggaac gcgttcaccg ccgggaaagt tatatttgca 600 15600 DNA Norwalk-like Virus (GII) 15 ctgaaacaat gataccacac tcccaaagacccatacaact aatgtctttg ctgggcgagg 60 ccgcactcca cggcccagca ttctacagcaaaattagcaa gctagtcatt gcagaactga 120 aggaaggtgg catggatttt tacgtgcccagacaagagcc aatgttcaga tggatgagat 180 tctcagatct gagcacgtgg gagggcgatcgcaatctggc tcccagcttt gtgaatgaag 240 atggcgtcga atgacgccaa cccatctgatgggtccgcag ccaacctcgt cccagaggtc 300 aataatgagg ttatggctct ggagcccgttgttggtgccg ctattgcggc acctgtggcg 360 ggccaacaaa acgtaattga cccctggattagaaacaatt ttgtacaagc ccctggtgga 420 gagttcacag tgtcccctag aaacgctccaggtgagatac tgtggagcgc gcccttgggc 480 cctgatctga acccctatct ttctcatttgtccagaatgt acaatggtta tgcaggtggt 540 tttgaagtgc aagtaatcct cgcggggaatgcgttcaccg ccgggaaagt catatttgca 600 16 600 DNA Norwalk-like Virus (GII)16 ttgagtcaat gatcccacac tctcagaggc ccatacagct tatgtcactc ttaggtgaag 60cagcactgca tgaaccatca ttctacagca agatcagcaa gcttgtgata tctgaactga 120aagaaggtgg aatggatttt tacgtgccca ggcaagaacc catgttcaga tggatgagat 180tctcagattt gagcacgtgg gagggcgatc gcaatcttgc tcccagtctt gtgaatgaag 240atggcgtcga atgacgccgc tccatctact gatggtgcag ccggcctcgt gccagaaagt 300aataatgagg tcatggctct tgaacccgtg gctggcgccg ccttggcagc cccggtcacc 360ggtcaaacaa atataataga cccttggatt agagcaaatt ttgtccaggc ccctaatggt 420gaatttacag tttctccccg caatgcccct ggtgaagtgc tattgaatct agagttgggt 480ccagaattga atccttatct ggcacattta gcaagaatgt ataacgggta tgccggtggg 540atggaggtgc aggtcatgtt ggctgggaac gcgttcacag ccggcaaatt ggtcttcgcc 600 17600 DNA Norwalk-like Virus (GII) 17 ctgaaacaat gataccacat tcccaaaggcccatacagtt gatgtctctg ctaggtgaag 60 ctgcattgca cggtccagca ttctacagcaaaatcagtaa actagtcatt tcagagttga 120 aggaaggtgg catggacttt tacgtgcccaggcaagagcc gatgttcaga tggatgagat 180 tctcagacct gagcacgtgg gagggcgatcgcaatctggc tcccagtttt gtgaatgaag 240 atggcgtcga atgacgccgc cccatctaatgatggtgcag ccggtctcgt accagaggtc 300 aacaacgaga cgatggccct cgaaccggtggctggggctt ctatagccgc ccctctaacc 360 ggtcaaaata atgtgataga cccctggattagaatgaact ttgtccaagc cccaaatgga 420 gaattcacag tgtctccccg caattctcctggtgaaatct tgctaaattt ggaattaggc 480 cctgaattaa atccattctc agcacacctttcaagaatgt ataatggtta tgccggcggg 540 gttgaagtgc aggtactact cgctgggaacgcgttcacag cgggaaaact ggtgtttgca 600 18 600 DNA Norwalk-like Virus (GII)18 gtgaaaccat gataccacat gcgcagagac ccgtgcagct catggcacta ctgggagagt 60cctccctaca tggaccctca ttttacagca aggtcagcaa gctggttata tctgaactta 120aggagggagg aatggatttt tatgtgccca gacaagagtc aatgttcagg tggatgaggt 180tctcagatct aagcacatgg gagggcgatc gcaatctggc ccccagtttt gtgaatgaag 240atggcgtcga atgacgctgc tccatctaat gatggtgccg cctgcctcgt cccagagatc 300aacaatgagg caatggcgct agagccagtg gccggttcag cgatagcagc tcccctcact 360ggccagcaaa atataattga tccctggatt atgaataatt ttgtacaagc acctggtggt 420gagtttacag tgtcacccag gaattcccct ggtgaagtgc ttcttaactt ggagttaggt 480ccagaaataa atccttattt ggctcatctc gctagaatgt acaatggtta tgcaggtgga 540tttgaagtgc aagtggtcct agctggaaat gcgtttacag caggaaagat tatctttgca 600 19600 DNA Norwalk-like Virus (GII) 19 gtgaaaccat gataccacat acgcagagacccgtgcagct catggcactg ctgggagaat 60 cctccctaca tggaccctca ttttacagcaaggttagcaa gctggttata tctgaactta 120 aggagggagg aatggacttt tatgtgcccagacaagagtc aatgttcagg tggatgaggt 180 tctcagatct aagcacatgg gagggcgatcgcaatctggc tcccagtttt gtgaatgaag 240 atggcgtcga atgacgctgc tccatctaatgatggtgccg ccggcctcgt cccagagatc 300 aacaatgagg caatggcgct agatccagtggcgggtgcag cgatagcagc acccctcact 360 ggccagcaaa atataattga tccctggattatgaataact ttgtgcaagc acctggtggt 420 gagtttacag tgtcacctag gaattcccctggtgaagtgc ttcttaattt ggaattaggt 480 ccagaaataa atccctattt ggctcatcttgctagaatgt acaatggtta tgcaggtggg 540 tttgaagtgc aagtggtcct ggctggaaatgcgtttacag cagcaaaggt gatctttgca 600 20 600 DNA Norwalk-like Virus (GII)20 ctgaaacaat gatatccaca ctcccgaaga cccatacgct aatgtcactg ttaggtgaag 60cagcactgca tggaccatca ttctacagta agattattaa gctagttatt gcagagctga 120aggaaggtgg catggacttt tacgtgccta gacaagaacc aatgttccgg tggatgaggt 180tctcagactt gagcacgtgg gagggcgatc gcaatctggc tcccagcttt gtgaatgaag 240atggcgtcga atgacgccac tccatctaat gatggtgccg ccggcctcgt cccagagatc 300aacaatgagg caatggcgct agacccagtg gcgggtgcag cgatagcagc acccctcact 360ggtcagcaaa acataattga tccctggatt atgaataatt ttgtgcaagc acctggtggt 420gagtttacag tgtcccctag gaatccccct ggtgaagtgc ttcttaattt ggaattgggc 480ccagaaataa acccctattt ggcccatctt gctagaatgt ataatggtta tgcaggtgga 540tttgaagtgc aggtagtcct ggctgggaat gcgtttacag caggaaagat aatctttgca 600 21600 DNA Norwalk-like Virus (GII) 21 ctgaaacaat gatatccaca ctcccaaagacccatacact aatgtctttg ctgggcgagg 60 ccgcactcca cggcccagca ttctacagcaaaattagcaa gctagtcatt gcagaactga 120 aggaaggtgg catggatttt tacgtgcccagacaagagcc aatgttcaga tggatgagat 180 tctcagatct gagcacgtgg gagggcgatcgcaatctggc tcccagcttt gtgaatgaag 240 atggcgtcga atgacgccaa cccatctgatgggtccgcag ccaacctcgt cccagaggtc 300 aataatgagg ttatggctct ggagcccgttgttggtgccg ctattgcggc acctgtggcg 360 ggccaacaaa acgtaattga cccctggattagaaacaatt ttgtacaagc ccctggtgga 420 gagttcacag tgtcccctag aaacgctccaggtgagatac tgtggagcgc gcccttgggc 480 cctgatctga acccctatct ttctcatttgtccagaatgt acaatggtta tgcaggtggt 540 tttgaagtgc aagtaatcct cgcggggaacgcgttcaccg ccgggaaagt catatttgca 600 22 600 DNA Norwalk-like Virus (GII)22 gtgaaaccat gatatccact gtgctgaaga ccgagacgct catggcacta ctgggagaat 60cctccctaaa tggaccctca ttttacagca aggtcagcaa gctggttata tctgaactta 120aggagggagg aatggatttt tatgtgccca gacaagagtc aatgttcagg tggatgaggt 180tctcagatct aagcacatgg gagggcgatc gcaatctggc ctccagtttt gtgaatgaag 240atggcgtcga atcgcgctgc tccatctaat gatggtgccg cctgcctcgt cccagagatc 300aacaatgagg caatggcgct agagccagtg gcgggtgcag cgatagcagc gcccctcact 360ggccagcaaa atataattga tccctggatt atgaataatt ttgtgcaagc acctggtggt 420gagtttacag tgtcacccag gaattcccct ggtgaagtgc ttcttaattt ggaattaggt 480ccagaaataa atccttattt ggctcatctt gctagaatgt acaatggtta tgcaggtgga 540tttgaagtgc aagtggtcct ggctggaaat gcgtttacag cagcaaaaat tatctttgca 600 23600 DNA Norwalk-like Virus (GII) 23 ctgaaacaat gataccacac tcccaaagacccatacaatt gatgtctttg ctgggcgagg 60 ctgcactcca cggcccagca ttctacagcaaaatcagcaa gctggtcatt gcagagctga 120 aggaaggtgg catggatttt tacgtgcccagacaagagcc aatgttcaga tggatgaggt 180 tctcagatct gagcacgtgg gagggcgatcgcaatctggc tcccagtttt gtgaatgaag 240 atggcgtcga gtgacgccgc tccatctaatgatggtgcag ccggtcttgt accagaggct 300 aacaatgaga ccatggcact tgaaccggtggctggggctt caatagccgc cccactcacc 360 ggtcaaaaca atattataga cccctggattagattaaatt ttgtgcaggc tcccaatggg 420 gagttcacgg tttcaccccg caactcgcccggggaagtcc tattaaactt ggaattaggc 480 cccgaactaa atccatacct agcacacctttctagaatgt ataatggtta tgcaggtggg 540 gttgaggtgc aagtactact ggctgggaatgcgttcacag ctggaaaatt ggtgtttgcg 600 24 600 DNA Norwalk-like Virus (GII)24 acgagagcat ggttccccat tctcagcgag ccacacagct catggccctt cttggtgagg 60cctcattgca tggcccccag ttttacaaga aagttagcaa gatggtcatc aatgagatta 120agagtggtgg tctggaattt tatgtgccca gacaagaggc catgtttagg tggatgagat 180tctctgacct cagcacatgg gagggcgatc gcaatcttgc tcccgagggt gtgaatgaag 240atggcgtcga atgacgccgc tccatcgaat gatggtgctg ccaacctcgt accagaggcc 300aacaatgagg ttatggcact tgaaccggtg gtaggagcct caatcgcagc tcctgttgtc 360ggtcagcaaa atataattga cccctggatt agagaaaatt ttgtccaagc accacagggc 420gagtttactg tttcgccaag gaattcgcct ggtgagatgc ttttaaacct tgagttgggc 480ccagaactta acccctattt gagtcatttg tcccccatgt acaacggata tgctggtggc 540atgcaggttc aggtggtcct agctgggaat gcgttcacag ctgggaaaat catctttgcc 600 25600 DNA Norwalk-like Virus (GII) 25 acgagagcat ggttccccat tctcagcgagccacacagct catggccctt cttggtgagg 60 cctcattgca tggcccccag ttttacaagaaagttagcaa gatggtcatc aatgagatta 120 agagtggtgg tctggaattt tatgtgcccagacaagaggc catgtttagg tggatgagat 180 tctctgacct cagcacatgg gagggcgatcgcaatcttgc tcccgagggt gtgaatgaag 240 atggcgtcga atgacgccgc tccatcgaatgatggtgctg ccaacctcgt accagaggcc 300 aacaatgagg ttatggcact tgaaccggtggtaggagcct caatcgcagc tcctgttgtc 360 ggtcagcaaa atataattga cccctggattagagaaaatt ttgtccaagc accacagggc 420 gagtttactg tttcgccaag gaattcgcctggtgagatgc ttttaaacct tgagttgggc 480 ccagaactta acccctattt gagtcatttgtcccccatgt acaacggata tgctggtggc 540 atgcaggttc aggtggtcct agctgggaatgcgttcacag ctgggaaaat catctttgcc 600 26 600 DNA Norwalk-like Virus (GII)26 atgaaagcat ggtcccccac tcccaacggg ccacacaact catggccctt cttggtgaag 60cttcattaca cggaccccaa ttctacaaga aggtcagtaa gatggttatc agtgagatta 120agagtggtgg tctggaattt tatgtgccca gacaagaggc catgtttagg tggatgagat 180tctctgacct cagcacatgg gagggcgatc gcaatcttgc tcccgagagt gtgaatgaag 240atggcgtcga atgacgctgc tccatcgaat gatggtgctg ccaacctcgt accagaggcc 300aacaatgagg ttatggcact tgaaccggtg gtgggagcct caattgcagc tcctgtcgtc 360ggtcaacaaa atataattga cccctggatt agagaaaatt ttgttcaggc accacagggt 420gagtttactg tttcaccaag aaactcgcct ggtgagatgc ttttaaatct tgaattaggc 480ccagagctca atccttacct gagtcattta tcccgcatgt ataatggtta tgctggtggc 540atgcaggttc aggtggtcct agctgggaac gcgttcacag ctggtaaaat catctttgcc 600 27600 DNA Norwalk-like Virus (GII) 27 atgaaagcat ggtcccccac tcccaacgggccacacaact catggccctt cttggtgaag 60 cttcattaca cggaccccaa ttctacaagaaggtcagtaa gatggttatc agtgagatta 120 agagtggtgg tctggaattt tatgtgcccagacaagaggc catgtttagg tggatgagat 180 tctctgacct cagcacatgg gagggcgatcgcaatcttgc tcccgagagt gtgaatgaag 240 atggcgtcga atgacgctgc tccatcgaatgatggtgctg ccaacctcgt accagaggcc 300 aacaatgagg ttatggcact tgaaccggtggtgggagcct caattgcagc tcctgtcgtc 360 ggtcaacaaa atataattga cccctggattagagaaaatt ttgttcaggc accacagggt 420 gagtttactg tttcaccaag aaactcgcctggtgagatgc ttttaaatct tgaattaggc 480 ccagagctca atccttacct gagtcatttatcccgcatgt ataatggtta tgctggtggc 540 atgcaggttc aggtggtcct agctgggaacgcgttcacag ctggtaaaat catctttgcc 600 28 600 DNA Norwalk-like Virus (GII)28 gtgaaaccat gataccacat gcgcagagac ccgtgcagct catggcacta ctgggagaat 60cctccctgca tggaccctca ttctacagca aggtcagcaa gctggttata tctgaactta 120aggagggagg aatggatttt tatgtgccca gacaagagtc aatgtttagg tggatgaggt 180tctcagatct gagcacatgg gagggcgatc gcaatctggc tcccagtttt gtgaatgaag 240atggcgtcga atgacgctgc tccatctaat gatggtgccg ccggcctcgt cccagagatc 300aacaatgagg caatggcgct agagccagtg gcgggcgcag cgatagcagc gcccctcact 360ggccagcaaa atataattga tccctggatt atgaataatt ttgtgcaagc acctggtggt 420gagtttacag tgtcacctag gaattcccct ggtgaagtgc ttcttaattt ggaattaggt 480ccagaaataa acccctattt ggctcatctt gctagaatgt acaatggtta tgcaggtgga 540tttgaagtgc aagtggtcct agctggaaat gcgtttacgg caggaaaggt tatctttgca 600 29600 DNA Norwalk-like Virus (GII) 29 ttgaaagcat gattccccac tcccagagagcaacccagct aatggccctc cttggggaag 60 cctcgttgca tggtccccag ttttacaaaaaggtgagtaa aatggtcatc aatgagatca 120 agagtggtgg tctggagttt tacgtgcccagacaggaggc catgttcaga tggatgagat 180 tttcagacct cagcacgtgg gagggcgatcgcaatctggc tcccgagaat gtgaatgaag 240 atggcgtcga atgacgcagc tccatcgaatgatggcgcgg ctggcctcgt accagagatc 300 aaccatgagg tcatggccat agaacctgttgcaggggcct ctttagcagc ccctgtcgta 360 ggacaactca atataattgt cccctggattagaaataatt ttggacaagc ccctcgtgga 420 gagtttatag tgtaccctag aatcgctccaggtgaatttt tattagatct agagttaggc 480 cctgagttga acccctacct tgctcaccttgcacgcatgt ataatgggca tgcaggtggt 540 atggaggtgc agatagtgct tgctgggaatgcgttcacag cgggcaaaat cctgtttgca 600 30 600 DNA Norwalk-like Virus (GII)30 gtgaaaccat gataccacat gcgcagagac ccgtgcagct catggcacta ctgggagaat 60cctccctgca tggaccctca ttctacagca aggtcagcaa gctggttata tctgaactta 120aggagggagg aatggatttt tatgtgccca gacaagagtc aatgtttagg tggatgaggt 180tctcagatct aagcacatgg gagggcgatc gcaatctggc tcccagtttt gtgaatgaag 240atggcgtcga atgacgctgc tccatctaat gatggtgccg ccggcctcgt cccagagatc 300aacaatgagg caatggcgct agagccagtg gcgggcgcag cgatagcagc gcccctcact 360ggccagcaaa atataattga tccctggatt atgaataatt ttgtgcaagc acctggtggt 420gagtttacag tgtcacctag gaattcccct ggtgaagtgc ttcttaattt ggaattaggt 480ccagaaataa acccctattt ggctcatctt gctagaatgt acaatggtta tgcaggtgga 540tttgaagtgc aagtggtcct agctggaaat gcgtttacag caggaaaggt tatctttgca 600 31507 DNA Norwalk-like Virus (GII) 31 atgaaacaat gatacctcac tctcagaggcccatacaact catggcccta cttggtgaag 60 cctctctaca cggaccctct ttctacagcaaaattagcaa attggtcata actgaactca 120 aggaaggtgg aatggatttt tacgtgccaagacaagagcc tatgtttagg tggatgagat 180 tctctgactt gagcacgtgg gagggcgatcgcaatctggc tcccagtttt gtgaatgaag 240 atggcgtcga atgacgctgc tccatcaaatgatggtgccg ccggcctcgt gccagaaagt 300 aataatgagg caatggccct ggaacccgtggtgggggtgt ctttagccgc ccctgtcact 360 ggccaaacta atataataga cccctggattagaactaatt ttgtccaagc ccctaatggt 420 gaatttacag tttcccctag aaattcccctggagagatat tggtcaattt ggagttgggt 480 ccagaactga atccttatct ggcacat 50732 513 DNA Norwalk-like Virus (GII) 32 atgagacaat gattccacac ttccagagacccatacaact aatgtcattg ctgggggaag 60 cagcattgca tggaccagct ttttacaagaaagttagcaa attagttatc actgagctca 120 aagagggtgg gatggatttc tatgtaccaagacaggaacc catgttcaga tggatgagat 180 tctcagacct cagtacttgg gagggcgatcgcaatcttgc tcccgaaggt gtgaatgaag 240 atggcgtcga atgacgctgc tccatctaatgatggtgccg ccggcctcgt cccagagatc 300 aacaatgagg caatggcgct agacccagtggcgggtgcag caatagcagc accccttact 360 ggccagcaaa atataattga tccctggattatgaataact ttgtgcaagc acctggtggt 420 gagttcacag tgtcacctag aaattcccctggtgaagtgt tacttaattt ggaattgggt 480 ccagaaataa atccctactt ggcccatcttgct 513 33 505 DNA Norwalk-like Virus (GII) misc_feature (1)..(1) n isa, c, g, or t 33 ntgaaaccat gattccacan tcncagagac ccntncagct natgncantnctgggngant 60 cntcnttgca tggaccnncn ttttacaaca aggtcagcaa nntggttatctctgagctna 120 angagggngg aatggatttt tatgtgccca gacaagagnc catgtttagntggatgagnt 180 tctcagacct cagcacatgg gagggcgatc gcaatctngc tcccaanngtgtgaatgaag 240 atggcgtcga atgacgctgc tccatctaat gatggtgccg ccggcctcgtcccagagatc 300 aacaatgagg caatggcgct aganccagtg gcgggtgcag cnatagcagcgcccctcact 360 ggccagcaaa atataattga tccctggatt atgaataant ttgtgcaagcacctggtggt 420 gagttnacag tgtcacctag naattcncct ggtgaagtgc ttcttaatttggaattaggt 480 ccagaaatna atccctactg gctct 505 34 516 DNA Norwalk-likeVirus (GII) 34 atgagacaat gataccccac tctcaaagac ccatacagct tatggcactgcttggtgagg 60 cctcccttca cggaccctct ttctacagca agattagtaa attggtcataactgaactca 120 aagaaggtgg gatggatttt tacgtgccaa gacaggaacc tatgttcaggtggatgaggt 180 tttctgacct gagcacgtgg gagggcgatc gcaatctggc tcccagttttgtgaatgaag 240 atggcgtcga atgacgccgc tccatctaat gatggtgcag ccggtctcgtaccagaggtc 300 aacaacgaga caatggcact tgagccagtt gcgggagctt caatcgccgcccctttaact 360 ggtcaaaaca atgtaataga cccctggatt agattaaatt ttgtacaagcccccaatggt 420 gagttcacgg tttccccccg aaactcgcct ggtgaaattt tgttaaatttggagttaggg 480 cctgaattga atccatattt agcccattta gcaaga 516 35 506 DNANorwalk-like Virus (GII) 35 atgagacaat gataccccac tctcaaagac ccatacagcttatggcactg cttggtgagg 60 cctcccttca cggaccctct ttctacagca agattagtaaattggtcata actgaactca 120 aagaaggtgg gatggatttt tacgtgccaa gacaggaacctatgttcagg tggatgaggt 180 tttctgacct gagcacgtgg gagggcgatc gcaatctggctcccagtttt gtgaatgaag 240 atggcgtcga atgacgccgc tccatctaat gatggtgcagccggtctcgt accagaggtc 300 aacaacgaga caatggcact tgagccagtt gcgggagcttcaatcgccgc ccctttaact 360 ggtcaaaaca atgtaataga cccctggatt agattaaattttgtacaagc ccccaatggt 420 gagttcacgg tttccccccg aaactcgcct ggtgaaattttgttaaattt ggagttaggg 480 cctgaattga atccatattt agccca 506 36 511 DNANorwalk-like Virus (GII) 36 atgagacaat gattccacac tcccagaggc ccatacaactaatgtcattg ctgggggagg 60 cagcattaca cggaccagct ttctacaaga aagtcagtaaattagttatc actgagctca 120 aagagggtgg aatggatttt tacgtgccaa ggcaggaacctatgtttagg tggatgagat 180 tctctgacct cagcacttgg gagggcgatc gcaatcttgctcccgaaggt gtgaatgaag 240 atggcgtcga atgacgctac tccatcaaat gatggtgctgccggcctcgt gccagaaagt 300 aacaatgagg caatggctct ggaacccgtg gtgggggcgtctttagccgc ccctgtcact 360 ggccaaacta atataataga cccctggatt agaactaattttgtccaagc ccctaatggt 420 gaattcacag tttcccctag aaattcccct ggagagatattggtcaattt ggagttgggt 480 ccagaactga acccttatct ggcacagtta g 511 37 511DNA Norwalk-like Virus (GII) 37 atgagacaat gataccccac tctcaaagacccatacagct tatggcactg cttggtgagg 60 cctcccttca cggaccctct ttctacagcaagattagtaa attggtcata actgaactca 120 aagaaggtgg gatggatttt tacgtgccaagacaggaacc tatgttcagg tggatgaggt 180 tttctgacct gagcacgtgg gagggcgatcgcaatctggc tcccagtttt gtgaatgaag 240 atggcgtcga atgacgccgc tccatctaatgatggtgcag ccggtctcgt accagaggtc 300 aacaacgaga caatggcact tgagccagttgcgggagctt caatcgccgc ccctttaact 360 ggtcaaaaca atgtaataga cccctggattagattaaatt ttgtacaagc ccccaatggt 420 gagttcacgg tttccccccg aaactcgcctggtgaaattt tgttaaattt ggagttaggg 480 cctgaattga atccatattt agcccattta g511 38 515 DNA Norwalk-like Virus (GII) 38 atgagacaat gatacctcactctcagaggc caatacaact catgtctcta cttggtgaag 60 ccgccctaca cggcccatcattttacagca aaatcagcaa gctggttatc acagaactaa 120 aggaaggtgg catggatttttacgtgccca ggcaagagcc catgtttagg tggatgagat 180 tctcagactt gagcacgtgggagggcgatc gcaatctggc tcccagtttt gtgaatgaag 240 atggcgtcga atgacgccgctccatctact gatggtgcag ccggcctcgt gccagaaagt 300 aataatgagg tcatggctcttgaacccgtg gctggtgccg ctttggcagc cccggtcacc 360 ggccaaacaa atattatagacccttggatt agagcaaatt ttgtccaggc ccctaatggt 420 gaatttacag tttctccccgtaatgcccct ggtgaagtgc tattgaatct agagttgggt 480 ccagaattaa atccttatctggcacattta gcaag 515 39 508 DNA Norwalk-like Virus (GII) 39 ctgaaacaatgattccacat tcccaaagac ccatacaatt gatgtcccta ctgggagagg 60 ccgcactccacggcccagca ttctacagca aaattagcaa gttagttatt gcagagctaa 120 aagaaggtggcatggatttt tacgtgccca gacaagagcc aatgttcaga tggatgagat 180 tctcagatctgagcacgtgg gagggcgatc gcaatctggc tcccagtttt gtgaatgaag 240 atggcgtcgaatgacgccaa cccatctgat gggtccacag ccaacctcgt cccagaggtc 300 aacaatgaggttatggcttt ggagcccgtt gttggtgccg ctattgcggc acctgtagcg 360 ggccaacaaaatgtaattga cccctggatt agaaataatt ttgtacaagc ccctggtgga 420 gagtttacagtatcccctag aaacgctccg ggtgagatac tatggagcgc gcccttgggc 480 cctatttgaacccctacctt ctcttggc 508 40 512 DNA Norwalk-like Virus (GII) 40ctgaaacaat gattccacat tcccaaagac ccatacaatt gatgtcccta ctgggagagg 60ccgcactcca cggcccagca ttctacagca aaattagcaa gttagttatt gcagagctaa 120aagaaggtgg catggatttt tacgtgccca gacaagagcc aatgttcaga tggatgagat 180tctcagatct gagcacgtgg gagggcgatc gcaatctggc tcccagtttt gtgaatgaag 240atggcgtcga atgacgccaa cccatctgat gggtccacag ccaacctcgt cccagaggtc 300aacaatgagg ttatggcttt ggagcccgtt gttggtgccg ctattgcggc acctgtagcg 360ggccaacaaa atgtaattga cccctggatt agaaataatt ttgtacaagc ccctggtgga 420gagtttacag tatcccctag aaacgctccg ggtgagatac tatggagcgc gcccttgggc 480cctgatttga acccctacct ttctcacttg gc 512 41 417 DNA Norwalk-like Virus(GII) 41 aatcagcaag ctggtcattg cagagctgaa ggaaggtggc atggatttttacgtgcccag 60 acaagagcca atgttcagat ggatgaggtt ctcagatctg agcacgtgggagggcgatcg 120 caatctggct cccagttttg tgaatgaaga tggcgtcgag tgacgccgctccatctaatg 180 atggtgcagc cggtcttgta ccagaggcta acaatgagac catggcacttgaaccggtgg 240 ctggggcttc aatagccgcc ccactcaccg gtcaaaacaa tattatagacccctggatta 300 gattaaattt tgtgcaggct cccaatgggg agttcacggt ttcaccccgcaactcgcccg 360 gtgaagtcct attaaacttg gaattaggcc ccgaactaaa tccatacctagcacacc 417 42 421 DNA Norwalk-like Virus (GII) 42 aatcagcaag ctggtcatcacagaactaaa ggaaggtggc atggattttt acgtgcccag 60 gcaagagccc atgtttaggtggatgagatt ctcagacttg agcacgtggg agggcgatcg 120 caatctggct cccagttttgtgaatgaaga tggcgtcgaa tgacgccgct ccatctactg 180 atggtgcagc cggcctcgtgccagaaagta ataatgaggt catggctctt gaacccgtgg 240 ctggtgccgc tttggcagccccggtcaccg gccaaacaaa tattatagac ccttggatta 300 gagcaaattt tgtccaggcccctaatggtg aatttacagt ttctccccgt aatgcccctg 360 gtgaagtgct attgaatctagagttgggtc cagaattaaa tctttatctg gctcttttag 420 c 421 43 422 DNANorwalk-like Virus (GII) 43 ggtcagtaag atggttatca gtgagatcaa gagtggtggtctggagtttt atgtgcccag 60 acaagaggcc atgtttaggt ggatgagatt ctctgacctcagcacatggg agggcgatcg 120 caatcttgct cccgagagtg tgaatgaaga tggcgtcgaatgacgctgct ccatcgaatg 180 atggtgctgc caacctcgta ccagaggcca acaatgaggttatggcactt gaaccggtgg 240 tgggagcctc aattgcagct cctgtcgtcg gtcaacaaaatataattgac ccctggatta 300 gagaaaattt tgttcaggca ccacagggtg agtttactgtttcaccaaga aactcgcctg 360 gtgagatgct tttaaatctt gaattaggcc cagagctcaatccttacctg agtcatttat 420 cc 422 44 422 DNA Norwalk-like Virus (GII) 44aattagcaag ttagttattg cagagctaaa agaaggtggc atggattttt acgtgcccag 60acaagagcca atgttcagat ggatgagatt ctcagatctg aacacgtggg agggcgatcg 120caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgccaac ccatctgatg 180ggtccacagc caacctcgtc ccagaggtca acaatgaggt tatggctttg gagcccgttg 240ttggtgccgc tattgcggca cctgtagcgg gccaacaaaa tgtaattgac ccctggatta 300gaaataattt tgtacaagcc cctggtggag agtttacagt atcccctaga aacgctccgg 360gtgagatact atggagcgcg cccttgggcc ctgatttgaa cccctacctt tctcatttgg 420 cc422 45 422 DNA Norwalk-like Virus (GII) 45 aattagcaag ttggttattgcagagctaaa agaaggtggc atggattttt acgtgcccag 60 acaagagcca atgttcagatggatgagatt ctcagatctg agcacgtggg agggcgatcg 120 caatctggct cccagttttgtgaatgaaga tggcgtcgaa tgacgccaac ccatctgatg 180 ggtccacagc caacctcgtcccagaggtca acaatgaggt tatggctttg gagcccgttg 240 ttggtgccgc tattgcggcacctgtagcgg gccaacaaaa tgtaattgac ccctggatta 300 gaaataattt tgtacaagcccctggtggag agtttacagt atcccctaga aacgctccgg 360 gtgagatact atggagcgcgcccttgggcc ctgatttgaa cccctacctt tctcatttgg 420 cc 422 46 410 DNANorwalk-like Virus (GII) 46 aattagcaag ttagttattg cagagctaaa agaaggtggcatggattttt acgtgcccag 60 acaagagcca atgttcagat ggatgagatt ctcagatctgagcacgtggg agggcgatcg 120 caatctggct cccagttttg tgaatgaaga tggcgtcgaatgacgccaac ccatctgatg 180 ggtccacagc caacctcgtc ccagaggtca acaatgaggttatggctttg gagcccgttg 240 ttggtgccgc tattgcggca cctgtagcgg gccaacaaaatgtaattgac ccctggatta 300 gaaataattt tgtacaagcc cctggtggag agtttacagtatcccctaga aacgctccgg 360 gtgagatact atggagcgcg cccttgggcc ctgatttgaacccctacctt 410 47 422 DNA Norwalk-like Virus (GII) 47 ggtcagcaagctggttatat ctgaacttaa ggagggagga atggattttt atgtgcccag 60 acaagagtcaatgtttaggt ggatgaggtt ctcagatcta agcacatggg agggcgatcg 120 caatctggctcccagttttg tgaatgaaga tggcgtcgaa tgacgctgct ccatctaatg 180 atggtgccgccggcctcgtc ccagagatca acaatgaggc aatggcgcta gagccagtgg 240 cgggcgcagcgatagcagcg cccctcactg gccagcaaaa tataattgat ccctggatta 300 tgaataattttgtgcaagca cctggtggtg agtttacagt gtcacctagg aattcccctg 360 gtgaagtgcttcttaatttg gaattaggtc cagaaataaa cccctatttg gctcatcttg 420 ct 422 48 422DNA Norwalk-like Virus (GII) 48 ggtcagcaag ctggttatat ctgaacttaaggagggagga atggattttt atgtgcccag 60 acaagagtca atgtttaggt ggatgaggttctcagatcta agcacatggg agggcgatcg 120 caatctggct cccagttttg tgaatgaagatggcgtcgaa tgacgctgct ccatctaatg 180 atggtgccgc cggcctcgtc ccagagatcaacaatgaggc aatggcgcta gagccagtgg 240 cgggcgcagc gatagcagcg ccactcactggccagcaaaa tataattgat ccctggatta 300 tgaataattt tgtgcaagca cctggtggtgagtttacagt gtcacctagg aattcccctg 360 gtgaagtgct tcttaatttg gaattaggtccagaaataaa cccctatttg gctcatcttg 420 ct 422 49 422 DNA Norwalk-likeVirus (GII) 49 ggtcagcaag ctggttatat ctgaacttaa ggagggagga atggatttttatgtgcccag 60 acaagagtca atgtttaggt ggatgaggtt ctcagatcta agcacatgggagggcgatcg 120 caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgctgctccatctaatg 180 atggtgccgc cggcctcgtc ccagagatca acaatgaggc aatggcgctagagccagtgg 240 cgggcgcagc gatagcagcg cccctcactg gccagcaaaa tataattgatccctggatta 300 tgaataattt tgtgcaagca cctggtggtg agtttacagt gtcacctaggaattcccctg 360 gtgaagtgct tcttaatttg gaattaggtc cagaaataaa cccctatttggctcatcttg 420 ct 422 50 422 DNA Norwalk-like Virus (GII) 50 agttagcaaattagttatca ctgagctcaa agagggtggg atggatttct atgtaccaag 60 acaggaacccatgttcagat ggatgagatt ctcagacctc agtacttggg agggcgatcg 120 caatcttgctcccgaaggtg tgaatgaaga tggcgtcgaa tgacgctgct ccatctaatg 180 atggtgccgccggcctcgtc ccagagatca acaatgaggc aatggcgcta gacccagtgg 240 cgggtgcagcaatagcagca ccccttactg gccagcaaaa tataattgat ccctggatta 300 tgaataactttgtgcaagca cctggtggtg agttcatagt gtcacctaga aattctcctg 360 gtgaagtgttacttaatttg gaattgggtc cagaaataaa tccctacttg gcccatcttg 420 ct 422 51 422DNA Norwalk-like Virus (GII) 51 ggtcagcaag ctggttatat ctgaacttaaggagggagga atggattttt atgtgcccag 60 acaagagtca atgtttaggt ggatgaggttctcagatcta agcacatggg agggcgatcg 120 caatctggct cccagttttg tgaatgaagatggcgtcgaa tgacgctgct ccatctaatg 180 atggtgccgc cggcctcgtc ccagagatcaacaatgaggc aatggcgcta gacccagtgg 240 cgggtgcagc aatagcagca ccccttactggccagcaaaa tataattgat ccctggatta 300 tgaataactt tgtgcaagca cctggtggtgagttcacagt gtcacctaga aattctcctg 360 gtgaagtgtt acttaatttg gaattaggtccagaaataaa cccctatttg gctcatcttg 420 ct 422 52 422 DNA Norwalk-likeVirus (GII) 52 ggtcagcaag ctggttatat ctgaacttaa ggagggagga atggatttttatgtgcccag 60 acaagagtca atgtttaggt ggatgaggtt ctcagatcta agcacatgggagggcgatcg 120 caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgctgctccatctaatg 180 atggtgccgc cggcctcgtc ccagagatca acaatgaggc aatggcgctagagccagtgg 240 cgggcgcagc gatagcagcg cccctcactg gccagcaaaa tataattgatccctggatta 300 tgaataattt tgtgcaagca cctggtggtg agtttacagt gtcacctaggaattcccctg 360 gtgaagtgct tcttaatttg gaattaggtc cagaaataaa cccctatttggctcatcttg 420 ct 422 53 422 DNA Norwalk-like Virus (GII) 53 ggtcagcaagctagttatat ctgaacttaa ggagggagga atggattttt atgtgcccag 60 acaagagtcaatgtttaggt ggatgaggtt ctcagatcta agcacatggg agggcgatcg 120 caatctggctcccagttttg tgaatgaaga tggcgtcgaa tgacgctgct ccatctaatg 180 atggtgccgccggcctcgtc ccagagatca acaatgaggc aatggcgcta gagccagtgg 240 cgggcgcagcgatagcagcg cccctcactg gccagcaaaa tataattgat ccctggatta 300 tgaataattttgtgcaagca cctggtggtg agtttacagt gtcacctagg aattcccctg 360 gtgaagtgcttcttaatttg gaattaggtc cagaaataaa cccttatttg gctcatcttg 420 ct 422 54 418DNA Norwalk-like Virus (GII) 54 aatcagcaag ctggtcattg cagagctgaaggaaggtggc atggattttt acgtgcccag 60 gcaagagcct atgttcagat ggatgaggttttcagatctg agcacgtggg agggcgatcg 120 caatctggct cccagttttg tgaatgaagatggcgtcgaa tgacgccgct ccatctaatg 180 atggtgcagc cggcctcgtg ccagaaagtaacaatgaggt tatggctctt gaacctgttg 240 ctggggcatc tttagctgcc cctgtgactggtcaaactaa tataattgac ccatggatca 300 gaatgaattt tgttcaagcc ccaaatggagaattcactgt ttccccaaga aattcccctg 360 gagaagtact cctaaatttg gaattgggtcctgaattaaa cccttatctg gcacacct 418 55 418 DNA Norwalk-like Virus (GII)55 aatcagcaag ctggtcattg cagagctgaa ggaaggtggc atggattttt acgtgcccag 60gcaagagcct atgttcagat ggatgaggtt ttcagatctg agcacgtggg agggcgatcg 120caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgccgct ccatctaatg 180atggtgcagc cggcctcgtg ccagaaagta acaatgaggt tatggctctt gaacctgttg 240ctggggcatc tttagctgcc cctgtgactg gtcaaactaa tataattgac ccatggatca 300gaatgaattt tgttcaagcc ccaaatggag aattcactgt ttccccaaga aattcccctg 360gagaagtact cctaaatttg gaattgggtc ctgaattaaa cccttatctg gcacacct 418 56422 DNA Norwalk-like Virus (GII) 56 ggtgagtaaa atggtcatca atgagatcaagagtggtggt ctggagtttt acgtgcccag 60 acaggaggcc atgttcaggt ggatgagattttcagacctc agcacgtggg agggcgatcg 120 caatctggct cccgagaatg tgaatgaagatggcgtcgaa tgacgcagct ccatcgaatg 180 atggcgcggc tggcctcgta ccagagatcaaccatgaggt catggccata gaacctgttg 240 caggggcatc tttagcagcc cctgtcgtaggacaacttaa cataattgat ccctggatta 300 gaaataattt tgtgcaagcc cctgctggagaatttactgt ttcacccaga aatgctccag 360 gtgaattttt gttagattta gagttaggccctgaattgaa cccctacctt gctcaccttg 420 ca 422 57 419 DNA Norwalk-likeVirus (GII) 57 aatcagcaag ctggtcattg cagagctgaa ggaaggtggc atggatttttacgtgcccag 60 gcaagagcct atgttcagat ggatgaggtt ttcagatctg agcacgtgggagggcgatcg 120 caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgccgctccatctaatg 180 atggtgcagc cggcctcgtg ccagaaagta acaatgaggt tatggctcttgaacctgttg 240 ctggggcatc tttagcagcc cctgtgactg gtcaaactaa tataattgacccatggatca 300 gaatgaattt tgttcaagcc ccaaatggag aattcactgt ttccccaagaaattcccctg 360 gagaagtact cctaaatttg gaattgggtc ctgaattaaa cccttatctggcacacctt 419 58 417 DNA Norwalk-like Virus (GII) 58 aatcagcaagctggtcattg cagagctgaa ggaaggtggc atggattttt acgtgcccag 60 gcaagagcctatgttcagat ggatgaggtt ttcagatctg agcacgtggg agggcgatcg 120 caatctggctcccagttttg tgaatgaaga tggcgtcgaa tgacgccgct ccatctaatg 180 atggtgcagccggcctcgtg ccagaaagta acaatgaggt tatggctctt gaacctgttg 240 ctggggcatctttagcagcc cctgtgactg gtcaaactaa tataattgac ccatggatca 300 gaatgaattttgttcaagcc ccaaatggag aattcactgt ttccccaaga aattcccctg 360 gagaagtactcctaaatttg gaattgggtc ctgaattaaa cccttatctg gcacacc 417 59 422 DNANorwalk-like Virus (GII) 59 aatcagcaag ctggtcattg cagagctgaa ggaaggtggcatggattttt acgtgcccag 60 gcaagagcct atgttcagat ggatgaggtt ttcagatctgagcacgtggg agggcgatcg 120 caatctggct cccagttttg tgaatgaaga tggcgtcgaatgacgccgct ccatctaatg 180 atggtgcagc cggcctcgtg ccagaaagta acaatgaggttacggctctt gaacctgttg 240 ctggggcatc tttagctgcc cctgtgactg gtcaaactaatataattgac ctatggacca 300 gaatgaatct tgttcaagcc ccaaatggag aattcactgtttccccaaga aattcccctg 360 gagaagtact cctaaatttg gaattgggtc ctgaattaaacccttatctg gcacacctat 420 ct 422 60 422 DNA Norwalk-like Virus (GII) 60aatcagcaag ctggtcattg cagagctgaa ggaaggtggc atggattttt acgtgcccag 60gcaagagcct atgttcagat ggatgaggtt ttcagatctg agcacgtggg agggcgatcg 120caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgccgct ccatctaatg 180atggtgcagc cggcctcgtg ccagaaagta acaatgaggt tatggctctt gaacctgttg 240ctggggcatc tttagctgcc cctgtgactg gtcaaactaa tataattgac ccatggatcg 300gaatgaattt tgttcaagcc ccaaatggag aattcactgt ttccccaaga aattcccctg 360gagaagtact cctaaatttg gaattgggtc ctgaattaaa cccttatctg gcacacctat 420 ct422 61 422 DNA Norwalk-like Virus (GII) 61 aatcagcaag ctggtcattgcagagctgaa ggaaggtggc atggattttt acgtgcccag 60 gcaagagcct atgttcagatggatgaggtt ttcagatctg agcacgtggg agggcgatcg 120 caatctggct cccagttttgtgaatgaaga tggcgtcgaa tgacgccgct ccatctaatg 180 atggtgcagc cggcctcgtgccagaaagta acaatgaggt tatggctctt gaacctgttg 240 ctggggcatc tttagctgcccctgtgactg gtcaaactaa tataattgac ccatggatca 300 gaatgaattt tgttcaagccccaaatggag aattcactgt ttccccaaga aattcccctg 360 gagaagtact cctaaatttggaattgggtc ctgaattaaa cccttatctg gcacacctat 420 ct 422 62 416 DNANorwalk-like Virus (GII) 62 agatagcaaa ttagttatca ctgagctcaa agagggtgggatggatttct ttgtaccaag 60 acaggaaccc atgttcagat ggatgagatt ctcagacctcagtacttggg agggcgatcg 120 caatcttgct cccgaaggtg tgaatgaaga tggcgtcgaatgacgctgct ccatctaatg 180 atggtgccgc cggcctcgtc ccagagatca acaatgaggcaatggcgcta gacccagtgg 240 cgggtgcagc aatagcagca ccccttactg gccagcaaaatataattgat ccctggatta 300 tgaataactt tgtgcaagca cctggtggtg agttcacagtgtcacctaga aattctcctg 360 gtgaagtgtt acttaatttg gaattgggtc cagaaataaatccctacttg gcccat 416 63 418 DNA Norwalk-like Virus (GII) 63 ggacagtaagatggttatca gtgagatcaa gagtggtggt ctggagtttt ttgtgcccag 60 acaagaggccatgtttaggt ggatgagatt ctctgacctc agcacatggg agggcgatcg 120 caatcttgctcccgagagtg tgaatgaaga tggcgtcgaa tgacgctgct ccatcgaatg 180 atggtgctgccaacctcgta ccagaggcca acaatgaggt tatggcactt gaaccggtgg 240 tgggagcctcaattgcagct cctgtcgtcg gtcaacaaaa tataattgac ccctggatta 300 gagaaaattttgttcaggca ccacagggtg agtttactgt ttcaccaaga aactcgcctg 360 gtgagatgcttttaaatctt gaattaggcc cagagctcaa tccttacctg agtcattt 418 64 418 DNANorwalk-like Virus (GII) 64 ggacagtaag atggttatca gtgagatcaa gagtggtggtctggagtttt ttgtgcccag 60 acaagaggcc atgtttaggt ggatgagatt ctctgacctcagcacatggg agggcgatcg 120 caatcttgct cccgagagtg tgaatgaaga tggcgtcgaatgacgctgct ccatcgaatg 180 atggtgctgc caacctcgta ccagaggcca acaatgaggttatggcactt gaaccggtgg 240 tgggagcctc aattgcagct cctgtcgtcg gtcaacaaaatataattgac ccctggatta 300 gagaaaattt tgttcaggca ccacagggtg agtttactgtttcaccaaga aactcgcctg 360 gtgagatgct tttaaatctt gaattaggcc cagagctcaatccttacctg agtcattt 418 65 418 DNA Norwalk-like Virus (GII) 65ggacagtaag atggttatca gtgagatcaa gagtggtggt ctggagtttt ttgtgcccag 60acaagaggcc atgtttaggt ggatgagatt ctctgacctc agcacatggg agggcgatcg 120caatcttgct cccgagagtg tgaatgaaga tggcgtcgaa tgacgctgct ccatcgaatg 180atggtgctgc caacctcgta ccagaggcca acaatgaggt tatggcactt gaaccggtgg 240tgggagcctc aattgcagct cctgtcgtcg gtcaacaaaa tataattgac ccctggatta 300gagaaaattt tgttcaggca ccacagggtg agtttactgt ttcaccaaga aactcgcctg 360gtgagatgct tttaaatctt gaattaggcc cagagctcaa tccttacctg agtcattt 418 66418 DNA Norwalk-like Virus (GII) 66 aatcagcaag ctggtcatca cagaactaaaggaaggtggc atggattttt tcgtgcccag 60 gcaagagccc atgtttaggt ggatgagattctcagacttg agcacgtggg agggcgatcg 120 caatctggct cccagttttg tgaatgaagatggcgtcgaa tgacgccgct ccatctactg 180 atggtgcagc cggcctcgtg ccagaaagtaataatgaggt catggctctt gaacccgtgg 240 ctggtgccgc tttggcagcc ccggtcaccggccaaacaaa tattatagac ccttggatta 300 gagcaaattt tgtccaggcc cctaatggtgaatttacagt ttctccccgt aatgcccctg 360 gtgaagtgct attgaatcta gagttgggtccagaattaaa tccttatctg gcacattt 418 67 422 DNA Norwalk-like Virus (GII)67 gattagtaaa ttggtcataa ctgaactcaa agaaggtggg atggattttt tcgtgccaag 60gcaggaacct atgttcaggt ggatgaggtt ttctgacttg agcacgtggg agggcgatcg 120caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgccgct ccatctactg 180atggtgcagc cggcctcgta ccagaggtca acaacgagac aatggcactt gaaccagttg 240cgggagcttc aatcgctgcc cccttaactg gtcaaaacaa tgtaatagac ccctggatta 300gattaaattt tgtacaagcc cccaatggtg agttcacggt ttccccccga aactcgcctg 360gtgaaatttt gttaaatttg gagttaggac ctgaattgaa cccatattta gcccatttag 420 ca422 68 422 DNA Norwalk-like Virus (GII) 68 gattagtaaa ttggtcataactgaactcaa agaaggtggg atggattttt tcgtgccaag 60 gcaggaacct atgttcaggtggatgaggtt ttctgacttg agcacgtggg agggcgatcg 120 caatctggct cccagttttgtgaatgaaga tggcgtcgaa tgacgccgct ccatctactg 180 atggtgcagc cggcctcgtaccagaggtca acaacgagac aatggcactt gaaccagttg 240 cgggagcttc aatcgctgcccccttaactg gtcaaaacaa tgtaatagac ccctggacta 300 gattaaatct tgtacaagcccccaatggtg agttcacggt ttccccccga aactcgcctg 360 gtgaaatttt gttaaatttggagttaggac ctgaattgaa cccatattta gcccatttag 420 ca 422 69 422 DNANorwalk-like Virus (GII) 69 ggtcagtaag atggttatca gtgagatcaa gagtggtggtctggagtttt ttgtgcccag 60 acaagaggcc atgtttaggt ggatgagatt ctctgacctcagcacatggg agggcgatcg 120 caatcttgct cccgagagtg tgaatgaaga tggcgtcgaatgacgctgct ccatcgaatg 180 atggtgctgc caacctcgta ccagaggcca acaatgaggttatggcactt gaaccggtgg 240 tgggagcctc aattgcagct cctgtcgtcg gtcaacaaaatataattgac ccctggatta 300 gagaaaattc tgttcaggca ccacagggtg agtttactgtttcaccaaga aactcgcctg 360 gtgagatgct tttaaatctt gaattaggcc cagagctcaatccttacctg agtcatttat 420 cc 422 70 422 DNA Norwalk-like Virus (GII) 70gattagtaaa ttggtcataa ctgaactcaa agaaggtggg atggattttt tcgtgccaag 60gcaggaacct atgttcaggt ggatgaggtt ttctgacttg agcacgtggg agggcgatcg 120caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgccgct ccatctactg 180atggtgcagc cggcctcgta ccagaggtca acaacgagac aatggcactt gaaccagttg 240cgggagcttc aatcgctgcc cccttaactg gtcaaaacaa tgtaatagac ccctggatta 300gattaaattt tgtacaagcc cccaatggtg agttcacggt ttccccccga aactcgcctg 360gtgaaatttt gttaaatttg gagttaggac ctgaattgaa cccatattta gcccatttag 420 ca422 71 410 DNA Norwalk-like Virus (GII) 71 aattagcaag ttagttattgcagagctaaa agaaggtggc atggattttt tcgtgcccag 60 acaagagcca atgttcagatggatgagatt ctcagatctg agcacgtggg agggcgatcg 120 caatctggct cccagttttgtgaatgaaga tggcgtcgaa tgacgccaac ccatctgatg 180 ggtccacagc caacctcgtcccagaggtca acaatgaggt tatggctttg gagcccgttg 240 ttggtgccgc tattgcggcacctgtagcgg gccaacaaaa tgtaattgac ccctggatta 300 gaaataattt tgtacaagcccctggtggag agtttacagt atcccctaga aacgctccgg 360 gtgagatact atggagcgcgcccttgggcc ctgatttgaa cccctacctt 410 72 422 DNA Norwalk-like Virus (GII)72 attaagcaag ttagttattg cagagctaaa agaaggtggc atggattttt tcgtgcccag 60acaagagcca atgttcagat ggatgagatt ctcagatctg agcacgtggg agggcgatcg 120caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgccaac ccatctgatg 180ggtccacagc caacctcgtc ccagaggtca acaatgaggt tatggctttg gagcccgttg 240ttggtgccgc tattgcggca cctgtagcgg gccaacaaaa tgtaattgac ccctggatta 300gaaataattt tgtacaagcc cctggtggag agtttacagt atcccctaga aacgctccgg 360gtgagatact atggagcgcg cccttgggcc ctgatttgaa cccctacctt tctcatttgg 420 cc422 73 422 DNA Norwalk-like Virus (GII) 73 ggacagtaag atggttatcagtgagatcaa gagtggtggt ctggagtttt ttgtgcccag 60 acaagaggcc atgtttaggtggatgagatt ctctgacctc agcacatggg agggcgatcg 120 caatcttgct cccgagagtgtgaatgaaga tggcgtcgaa tgacgctgct ccatcgaatg 180 atggtgctgc caacctcgtaccagaggcca acaatgaggt tatggcactt gaaccggtgg 240 tgggagcctc aattgcagctcctgtcgtcg gtcaacaaaa tataattgac ccctggatta 300 gagaaaattt tgttcaggcaccacagggtg agtttactgt ttcaccaaga aactcgcctg 360 gtgagatgct tttaaatcttgaattaggcc cagagctcaa tccttacctg agtcatttat 420 cc 422 74 422 DNANorwalk-like Virus (GII) 74 aatcagcaag ctggttatca cagaactaaa ggaaggtggcatggattttt tcgtgcccag 60 gcaagagccc atgtttaggt ggatgagatt ctcagacttgagcacgtggg agggcgatcg 120 caatctggct cccagttttg tgaatgaaga tggcgtcgaatgacgccgct ccatctactg 180 atggtgcagc cggcctcgtg ccagaaagta ataatgaggtcatggctctt gaacccgtgg 240 ctggtgccgc tttggcagcc ccggtcaccg gccaaacaaatattatagac ccttggatta 300 gagcaaattt tgtccaggcc cctaatggtg aatttacagtttctccccgt aatgcccctg 360 gtgaagtgct attgaatcta gagttgggtc cagaattaaatccttatctg gcacatttag 420 ca 422 75 422 DNA Norwalk-like Virus (GII) 75aatcagcaag ctggttatca cagaactaaa ggaaggtggc atggattttt tcgtgcccag 60gcaagagccc atgtttaggt ggatgagatt ctcagacttg agcacgtggg agggcgatcg 120caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgccgct ccatctactg 180atggtgcagc cggcctcgtg ccagaaagta ataatgaggt catggctctt gaacccgtgg 240ctggtgccgc tttggcagcc ccggtcaccg gccaaacaaa tattatagac ccttggatta 300gagcaaattt tgtccaggcc cctaatggtg aatttacagt ttctccccgt aatgcccctg 360gtgaagtgct attgaatcta gagttgggtc cagaattaaa tccttatctg gcacatttag 420 ca422 76 422 DNA Norwalk-like Virus (GII) 76 ggtcagcaag ctggttatatctgaacttaa ggagggagga atggattttt ttgtgcccag 60 acaagagtca atgtttaggtggatgaggtt ctcagatcta agcacatggg agggcgatcg 120 caatctggct cccagttttgtgaatgaaga tggcgtcgaa tgacgctgct ccatctaatg 180 atggtgccgc cggcctcgtcccagagatca acaatgaggc aatggcgcta gagccagtgg 240 cgggtgcagc gatagcagcgcccctcactg gccagcaaaa tataattgat ccctggatta 300 tgaataattt tgtgcaagcacctggtggtg agtttacagt gtcacctagg aattctcctg 360 gtgaagtgct tctcaatttggaattaggtc cagaaataaa tccctatttg gctcatcttg 420 ct 422 77 422 DNANorwalk-like Virus (GII) 77 agttagcaaa ttagttatca ctgagctcaa agagggtgggatggatttct ttgtaccaag 60 acaggaaccc atgttcagat ggatgagatt ctcagacctcagtacttggg agggcgatcg 120 caatcttgct cccgaaggtg tgaatgaaga tggcgtcgaatgacgctgct ccatctaatg 180 atggtgccgc cggcctcgtc ccagagatca acaatgaggcaatggcgcta gacccagtgg 240 cgggtgcagc gatagcagcg cccctcactg gccagcaaaatataattgat ccctggatta 300 tgaataactt tgtgcaagca cctggtggtg agttcacagtgtcacctaga aattctcctg 360 gtgaagtgtt acttaatttg gaattgggtc cagaaataaatccctacttg gcccatcttg 420 ct 422 78 422 DNA Norwalk-like Virus (GII) 78aatcagcaag ctggttatca cagaactaaa ggaaggtggc atggattttt tcgtgcccag 60gcaagagccc atgtttaggt ggatgagatt ctcagacttg agcacgtggg agggcgatcg 120caatctggct cccagttttg tgaatgaaga tggcgtcgaa tgacgccgct ccatctactg 180atggtgcagc cggcctcgtg ccagaaagta ataatgaggt catggctctt gaacccgtgg 240ctggtgccgc tttggcagcc ccggtcaccg gccaaacaaa tattatagac ccttggatta 300gagcaaattt tgtccaggcc cctaatggtg aatttacagt ttctccccgt aatgcccctg 360gtgaagtgct attgaatcta gagttgggtc cagaattaaa tccttatctg gcacatttag 420 ca422 79 422 DNA Norwalk-like Virus (GII) 79 aatcagcaag ctggtcatcacagaactaaa ggaaggtggc atggattttt tcgtgcccag 60 gcaagagccc atgtttaggtggatgagatt ctcagacttg agcacgtggg agggcgatcg 120 caatctggct cccagttttgtgaatgaaga tggcgtcgaa tgacgccgct ccatctactg 180 atggtgcagc cggcctcgtgccagaaagta ataatgaggt catggctctt gaacccgtgg 240 ctggtgccgc tttggcagccccggtcaccg gccaaacaaa tattatagac ccttggatta 300 gagcaaattt tgtccaggcccctaatggtg aatttacagt ttctccccgt aatgcccctg 360 gtgaagtgct attgaatctagagttgggtc cagaattaaa tccttatctg gcacatttag 420 ca 422 80 422 DNANorwalk-like Virus (GII) 80 81 296 DNA Norwalk-like Virus (GII) 81tgctcccgaa ggtgtgaatg aagatggcgt cgaatgacgc tgctccatct aatgatggtg 60ccgccggcct cgtcccagag atcaacaatg aggcaatggc gctagaccca gtggcgggtg 120cagcaatagc agcacccctt actggccagc aaaatataat tgatccctgg attatgaata 180actttgtgca agcacctggt ggtgagttca cagtgtcacc tagaaattct cctggtgaag 240tgttacttaa tttggaattg ggtccagaaa taaatcccta cttggcccat cttgct 296 82 296DNA Norwalk-like Virus (GII) 82 tgctcccgaa ggtgtgaatg aagatggcgtcgaatgacgc tgctccatct aatgatggtg 60 ccgccggcct cgtcccagag atcaacaatgaggcaatggc gctagaccca gtggcgggtg 120 cagcaatagc agcacccctt actggccagcaaaatataat tgatccctgg attatgaata 180 actttgtgca agcacctggt ggtgagttcacagtgtcacc tagaaattct cctggtgaag 240 tgttacttaa tttggaattg ggtccagaaataaatcccta cttggcccat cttgct 296 83 296 DNA Norwalk-like Virus (GII) 83tgctcccgaa ggtgtgaatg aagatggcgt cgaatgacgc tgctccatct aatgatggtg 60ccgccggcct cgtcccagag atcaacaatg aggcaatggc gctagaccca gtggcgggtg 120cagcaatagc agcacccctt actggccagc aaaatataat tgatccctgg attatgaata 180actttgtgca agcacctggt ggtgagttca cagtgtcacc tagaaattct cctggtgaag 240tgttacttaa tttggaattg ggtccagaaa taaatcccta cttggcccat cttgct 296 84 403DNA Norwalk-like Virus (GII) 84 tgggagggcg atcgcaatct ggctcccagttttgtgaatg aagatggcgt cgaatgacgc 60 tgctccatct aatgatggtg ccgccggcctcgtcccagag atcaacaatg aggcaatggc 120 gctagaccca gtggcgggtg cagcgatagcagcgcccctc actggccagc aaaatataat 180 tgatccctgg attatgaata actttgtgcaagcacctggt ggtgagttca cagtgtcacc 240 taggaattcc cctggtgaag tgcttcttaatttggagtta ggtccagaaa taaaccccta 300 tttggctcac cttgctagaa tgtacaatggttatgcaggt ggatttgaag tgcaagtggt 360 cctagctgga aatgcgttta cagcaggaaaggttatcttt gca 403 85 384 DNA Norwalk-like Virus (GII) 85 tggctcccagttttgtgaat gaagatggcg tcgaatgacg ccgctccatc tactgatggt 60 gcagccggcctcgtgccaga aagtaataat gaggtcatgg ctcttgagcc cgtggctggt 120 gctgccttggcagccccggt caccggtcaa acaaatatta tagacccttg gattagagca 180 aattttgtccaggcccctaa tggtgaattt acagtttctc cccgtaatgc ccctggtgaa 240 gtgctattaaatctagaatt gggtccagaa ttaaatcctt atctggcaca tttagcaaga 300 atgtacaacgggtatgccgg tgggatggag gtgcaggtca tgctagctgg gaacgcgttc 360 acagctggcaaattggtctt cgct 384 86 366 DNA Norwalk-like Virus (GII) 86 atgaagatggcgtcgaatga cgccgctcca tctaatgatg gtgcagccgg tcttgtacca 60 gaggctaacaatgagaccat ggcacttgaa ccggtggctg gggcttcaat agccgcccca 120 ctcaccggccaaaacaatat tatagacccc tggattagat taaattttgt gcaggctccc 180 aatggagagttcacggtttc accccgcaac tcacccgggg aagtcctatt aaatttggaa 240 ttaggccccgaactaaatcc atacctagca cacctttcta gaatgtataa tggttatgca 300 ggtggggttgaggtgcaagt actactggct gggaatgcgt tcacagctgg aaaattggtg 360 tttgcc 366 87366 DNA Norwalk-like Virus (GII) 87 atgaagatgg cgtcgaatga cgccaacccatctgatgggt ccacagccaa cctcgtccca 60 gaggtcaaca atgaggttat ggctttggagcccgttgttg gtgccgctat tgcggcacct 120 gtagcgggcc aacaaaatgt aattgacccctggattagaa ataattttgt acaagcccct 180 ggtggagagt ttacagtatc ccctagaaacgctccgggtg agatactatg gagcgcgccc 240 ttaggccctg atttgaaccc ctacctttctcatttggcca gaatgtacaa tggttatgca 300 ggtggttttg aagtgcaggt aatcctcgcggggaacgcgt tcaccgccgg gaaaatcata 360 tttgca 366 88 366 DNA Norwalk-likeVirus (GII) 88 atgaagatgg cgtcgaatga cgccaaccca tctgatgggt ccacagccaacctcgtccca 60 gaggtcaaca atgaggttat ggctttggag cccgttgtcg gtgccgctattgcggcacct 120 gtagcgggcc aacaaaatgt aattgacccc tggattagaa ataattttgtacaagcccct 180 ggtggagagt ttacagtatc ccctagaaac gctccgggtg agatactatggagcgcgccc 240 ttgggccctg atttgaaccc ctacctttct catttggcca gaatgtacaatggttatgca 300 ggtggttttg aagtgcaggt aatcctcgcg gggaacgcgt tcaccgccgggaaaatcata 360 tttgca 366 89 366 DNA Norwalk-like Virus (GII) 89atgaagatgg cgtcgaatga cgccaaccca tctgatgggt ccacagccaa cctcgtccca 60gaggtcaaca atgaggttat ggctttggag cccgttgttg gtgccgctat tgcggcacct 120gtagcgggcc aacaaaatgt aattgacccc tggattagaa ataattttgt acaagcccct 180ggtggagagt ttacagtatc ccctagaaac gctccgggtg agatactatg gagcgcgccc 240ttgggccctg atttgaaccc ttacctttct catttggcca gaatgtacaa tggttatgca 300ggtggttttg aagtgcaggt aatcctcgcg gggaacgcgt tcaccgccgg gaaaatcata 360tttgca 366 90 366 DNA Norwalk-like Virus (GII) 90 atgaagatgg cgtcgaatgacgccaaccca tctgatgggt ccacagccaa cctcgtccca 60 gaggtcaaca atgaggttatggctttggag cccgttgttg gtgccgctat tgcggcacct 120 gtagcgggcc aacaaaatgtaattgacccc tggattagaa ataattttgt acaagcccct 180 ggtggagagt ttacagtatcccctagaaac gctccgggtg agatactatg gagcgcgccc 240 ttaggccctg atttgaacccctacctttct catttggcca gaatgtacaa tggttatgca 300 ggtggttttg aagtgcaggtaatcctcgcg gggaacgcgt tcaccgccgg gaaaatcata 360 tttgca 366 91 366 DNANorwalk-like Virus (GII) 91 atgaagatgg cgtcgaatga cgccaaccca tctgatgggtccacagccaa cctcgtccca 60 gaggtcaaca atgaggttat ggctttggag cccgttgttggtgccgctat tgcggcacct 120 gtagcgggcc aacaaaatgt aattgacccc tggattagaaataattttgt acaagcccct 180 ggtggagagt ttacagtatc ccctagaaac gctccgggtgagatactatg gagcgcgccc 240 ttaggccctg atttgaaccc ctacctttct catttggccagaatgtacaa tggttatgca 300 ggtggttttg aagtgcaggt aatcctcgcg gggaacgcgttcaccgccgg gaaaatcata 360 tttgca 366 92 366 DNA Norwalk-like Virus (GII)92 atgaagatgg cgtcgaatga cgccaaccca tctgatgggt ccacagccaa cctcgtccca 60gaggtcaaca atgaggttat ggctttggag cccgttgttg gtgccgctat tgcggcacct 120gtagcgggcc aacaaaatgt aattgacccc tggattagaa ataattttgt gcaagcccct 180ggtggagagt ttacagtatc ccctagaaac gctccgggtg agatactatg gagcgcgccc 240ttaggccctg atttgaaccc ctacctttct catttggcca gaatgtacaa tggttatgca 300ggtggttttg aagtgcaagt aatcctcgcg gggaacgcgt tcaccgccgg gaaaatcata 360tttgca 366 93 366 DNA Norwalk-like Virus (GII) 93 atgaagatgg cgtcgaatgacgccaaccca tctgatgggt caacagccaa cctcgtccca 60 gaggtcaaca atgaggttatggctttggag cccgttgttg gtgccgctat tgcggcacct 120 gtagcgggcc aacaaaatgtaattgacccc tggattagaa ataattttgt acaagcccct 180 ggtggagagt ttacagtatcccctagaaac gctccgggtg agatactatg gagcgcgccc 240 ttaggccctg atttgaacccctacctttct catttggcca gaatgtacaa tggttatgca 300 ggtggttttg aagtgcaggtaatcctcgcg gggaacgcgt tcaccgccgg gaaaatcata 360 tttgca 366 94 366 DNANorwalk-like Virus (GII) 94 atgaagatgg cgtcgaatga cgccaaccca tctgatgggtccacagccaa cctcgtccca 60 gaggtcaaca atgaggttat ggctttggag cccgttgttggtgccgctat tgcggcacct 120 gtagcgggcc aacaaaatgt aattgacccc tggattagaaataattttgt acaagcccct 180 ggtggagagt ttacagtatc ccctagaaac gctccgggtgagatactatg gagcgcgccc 240 ttaggccctg atttgaaccc ctacctttct catttggccagaatgtacaa tggttatgca 300 ggtggttttg aagtgcaggt aatcctcgcg gggaacgcgttcaccgccgg gaaaatcata 360 tttgca 366 95 366 DNA Norwalk-like Virus (GII)95 atgaagatgg cgtcgaatga cgccaaccca tctgatgggt ccacagccaa cctcgtccca 60gaggtcaaca atgaggttat ggctttggag cccgttgttg gtgccgctat tgcggcacct 120gtagcgggcc aacaaaatgt aattgacccc tggattagaa ataattttgt acaagcccct 180ggtggagagt ttacagtgtc ccctagaaac gctccgggtg agatactatg gagcgcgccc 240ttaggccctg acttgaaccc ctacctttct catttggcca gaatgtacaa tggttatgca 300ggtggttttg aagtgcaggt aatcctcgcg gggaacgcgt tcaccgccgg gaaaatcata 360tttgca 366 96 366 DNA Norwalk-like Virus (GII) 96 atgaagatgg cgtcgaatgacgccaaccca tctgatgggt ccacagccaa cctcgtccca 60 gaggtcaaca atgaggttatggctttggag cccgttgttg gtgccgccat tgcggcacct 120 gtagcgggcc aacaaaatgtaattgacccc tggattagaa ataattttgt acaagcccct 180 ggtggagagt ttacagtgtcccctaggaac gctccgggtg agatactatg gagcgcgccc 240 ttgggccctg atttgaacccctacctttct cacttggcca gaatgtataa tggttatgca 300 ggtggttttg aagtgcaggtaatcctcgcg gggaacgcgt tcaccgccgg gaaaatcata 360 tttgca 366 97 366 DNANorwalk-like Virus (GII) 97 atgaagatgg cgtcgaatga cgccaaccca tctgatgggtccacagccaa cctcgtccca 60 gaagtcaaca atgaggttat ggctttggag cccgttgttggtgccgctat tgcggcacct 120 gtagcgggcc aacaaaatgt aattgacccc tggattagaaataattttgt acaagcccct 180 ggtggagagt ttacagtgtc ccctagaaac gctccgggtgaaatactatg gagcgcgccc 240 ttgggccctg atttgaaccc ctacctttcc catttggccagaatgtacaa tggttatgca 300 ggtggttttg aagtgcaggt aatcctcgcg gggaacgcgttcaccgccgg gaaaatcata 360 tttgca 366 98 366 DNA Norwalk-like Virus (GII)98 atgaagatgg cgtcgagtga cgctgctcca tctgcggatg gtgcgggcaa cctcgtccca 60gagagtcaac aagaggtatt gcccctcgcc cccgttgcgg gcgctgcact agcggcaccc 120gtagtggggc agacaaacat aattgacccc tggattaaag aaaattttgt tcaagccccc 180cagggtgagt ttacagtctc acctaaaaat tctcctggtg aaattttagt caatttggaa 240ttgggaccca aactcaaccc ctatctggat cacctctcac gcatgtacaa ttcatatgct 300ggtggtatag atgttatggt ggtgttggcg ggtaacgcct tcacagccgg taaggttcta 360atagca 366 99 366 DNA Norwalk-like Virus (GII) 99 atgaagatgg cgtcgaatgacgcagctcca tcgaatgatg gcgcggctgg cctcgtacca 60 gagatcaacc atgaggtcatggccatagaa cctgttgcag gggcctcttt agcagcccct 120 gtcgtaggac aacttaatataattgatccc tggattagaa ataattttgt acaagcccct 180 gctggagaat ttactgtttcacctagaaat gctccgggtg aatttttgtt agatttagag 240 ctaggccctg aattaaacccctatcttgct caccttgcac gcatgtataa tgggcatgct 300 ggtggtatgg aggtgcagatagtgcttgct gggaatgcgt tcacagcggg caaaatcctg 360 tttgca 366 100 366 DNANorwalk-like Virus (GII) 100 atgaagatgg cgtcgaatga cgccaaccca tctgatgggtccacagccaa cctcgtccca 60 gaggtcaaca atgaggttat ggctttggaa cccgttgttggtgccgctat tgcggcacct 120 gtagcgggcc aacaaaatgt aattgacccc tggattagaaataattttgt acaagcccct 180 ggtggagagt ttacagtatc ccctagaaac gctccgggtgagatactatg gagcgcgccc 240 ttgggccctg atttgaaccc ctacctttct catttggccagaatgtacaa tggttatgca 300 ggtggttttg aagtgcaggt aatcctcgcg gggaacgcgttcaccgccgg gaaaatcata 360 tttgca 366 101 366 DNA Norwalk-like Virus(GII) misc_feature (61)..(61) n is a, c, g, or t 101 atgaaratggcgtcgaatga cgccgctcca tctaatgatg gtgcagccgg tctcgtacca 60 naggtcaacaacgaracrat ggcactcgaa ccggtggctg gggcttccat agccgcccct 120 ctaaccggtcaaaacaatgt gatagacccc tggattagaa tgaactttgt tcaagcccca 180 aatggtgaatttacagtgtc tccccgtaac tctcctggtg aaattctgtt aaatttagaa 240 ttaggtcctgaattaaatcc attcttagca cacctttcaa ggatgtwtaa tggttatgct 300 ggtggggttgaaatgcaggt gctacttgct gggaacgcgt tcacagcagg aaaactactg 360 tttgca 366102 366 DNA Norwalk-like Virus (GII) 102 atgaagatgg cgtcgaatgacgctactcca tcaaatgatg gtgccgccgg cctcgtgcca 60 gaaagtaata atgaggcaatggctctggaa cccgtggctg gggcgtcttt agccgcccct 120 gtcactggcc aaactaatataatagacccc tggattagaa ctaattttgt ccaagcccct 180 aatggtgaat tcacagtttcccctaaaaat tcccctggag agatattggt caatttggag 240 ttgggtccag aactgaacccttatctggca catttagcta ggatgtacaa tggttatgcg 300 ggtggtatgg aggtgcaagtgatgcttgcg gggaacgcgt tcactgctgg caagatcatc 360 tttgcc 366 103 366 DNANorwalk-like Virus (GII) 103 atgaagatgg cgtcgaatga cgcagctcca tctaatgatggtgcagcagg cctcgtacca 60 garatcaaca atgaggtcat gccccttgag cccgtggctggtgcatcgct ggcgacacca 120 gttgttgggc aacaaaacat aattgatccc tggataagaaataattttgt gcaagcccct 180 gcaggtgagt ttacagtctc ccctaggaat tcccccggtgaaatcctgct tgatttagag 240 ttgggaccag aattgaaccc ctaccttgct catttggctcgtatgtataa tggacacgct 300 ggtggcatgg aagtgcaaat tgtgttggct gggaatgcgttcacagctgg caagatcgta 360 tttgct 366 104 366 DNA Norwalk-like Virus(GII) 104 atgaagatgg cgtcgaatga cgctgctcca tctaatgatg gtgccgccggcctcgctcca 60 gagatcaaca atgaggcaat ggcgctagag ccagtggcgg gtgcagcgatagcagcgccc 120 ctcactggcc agcaaaatat aattgatccc tggattatga ataattttgtgcaagcacct 180 ggtggtgagt ttacagtgtc acctaggaat tctcctggtg aagtgcttcttaatttggaa 240 ttaggtccag aaataaatcc ctatttggct catcttgcta gaatgtacaatggttatgca 300 ggtggatttg aagtgcaagt ggtcctagct ggaaatgcgc ttacagcaggaaagggtatc 360 tttgca 366 105 366 DNA Norwalk-like Virus (GII) 105atgaagatgg cgtcgaatsa cgccaaccca tctgatgggt ccacagccaa cctcgtcyca 60gaggtcaaca atgaggttat ggctttggag cccgttgttg gtgccgctat tgcagcacct 120gtagcgggcc aacaaaatgt aattgacccc tggattagaa ataattttgt acaagcccct 180ggtggagagt ttacagtatc ccctaaaaac gctccgggtg agatactatg gagcgcgccc 240ttgggccctg atttgaaccc ctacctttct catttggcca gaatgtacaa tggttatgca 300ggtggttttg aagtgcaggt aatcctcgcg gggaacgcgt tcaccgccgg gaaaatcata 360tttgca 366 106 366 DNA Norwalk-like Virus (GII) 106 atgaagatggcgtcgaatga cgctgctcca tctaatgatg gtgccgccgg cctcgtccca 60 gagatcaacaatgaggcaat ggcgctagag ccagtggcgg gtgcagcgat agcagcgccc 120 ctcactggccagcaaaatat aattgatccc tggattatga ataattttgc gcaagcacct 180 ggtggtgagtttacagtgtc acctaggaac tcccctggtg aagtgcttct taatttggaa 240 ttaggtccagaaataaatcc ctatttggct catcttgcta gaatgtacaa tggttatgca 300 ggtggatttgaagtgcaagt ggtcctagct ggaaatgcgt ttacagcagg aaaggttatc 360 tttgca 366107 366 DNA Norwalk-like Virus (GII) 107 atgaagatgg cgtcgagtgacgccgctcca tctaatgatg gtgcagccgg tctcgtacca 60 gaggctaacg wtgagaccatggcacttgaa ccggtggctg gggcttcaat agccgcccca 120 ctcaccggcc aaaacaatattatagacccc tggattarat taaactttgt gcaggctccc 180 aatggaragt tcacggtttcaccccgcaac tcacctgggg aagtcctact aaatttggaa 240 ttaggccccg aactaaatccatatctggca cacctttcta aaatgtataa tggttatgca 300 ggtggggttg aggtgcaagtactactggct gggwacgcgt tcacagctgg aaaattggtg 360 ttcgcc 366 108 366 DNANorwalk-like Virus (GII) 108 atgaagatgg cgtcgaatga cgccaaccca tctgatgggtccacagccaa cctcgtccca 60 gaggtcaaca atgaggttat ggctttggag cccgttgttggtgccgctat cgcggcacct 120 gtagcgggcc aacaaaatgt aattgacccc tggattagaagtaattttgt acaagcccct 180 ggtggagagt ttacagtatc ccctagaaac gctccaggtgagatactatg gagcgcgccc 240 ttgggccctg atttgaaccc ctacctttct catttggccagaatgtacaa tggttatgca 300 ggtggtcttg aagtgcaggt agtcctcgcg ggaaacgcgttcaccgccgg gaaaatcata 360 tttgca 366 109 366 DNA Norwalk-like Virus(GII) 109 atgaaratgg cgtcgaatga cgctgctcca tcgaatgatg gtgctgccaacctcgtacca 60 gaggccagca atgaggttat ggcacttgaa ccggtggtgg gagcctcaatcgcagctcct 120 gttgtcggtc agcaaaatat aattgacccc tggattagag aaaattttgtccaagcacca 180 cagggcgagt tcactgtttc accaaggaat tcgcctggtg agatgctcttaaaccttgag 240 ttgggcccag aacttaatcc ctatttgagt catttgtccc gcatgtacaacggatatgct 300 ggtggcatgc aggttcaggt ggtcctagct gggaatgcgt kcacagctgggaaaatcatc 360 tttgcc 366 110 366 DNA Norwalk-like Virus (GII) 110atgaagatgg cgtcgaatga cgccgctcca tctactgatg gtgcagccgg cctcgtgcca 60gaaagtaata atgaggtcat ggctcttgag cccgtggctg gtgctgcctt ggcagccccg 120gtcaccggtc aaacaaatat tatagaccct tggattagag caaattttgt ccaggcccct 180aatggcgaat ttacagtttc tccccgtaat gcccctggtg aagtgctatt aaatctagaa 240ttgggtccag aattaaatcc ttatctggca catttagcaa gaatgtacaa cgggtatgcc 300ggtgggatgg aggtgcaggt catgctagct gggaacgcgt tcacagctgg caaattggtc 360ttcgct 366 111 345 DNA Norwalk-like Virus (GII) 111 gctgctccgtctaatgatgg tgctgccggc ctcgttccag agatcaacaa tgaggcaatg 60 gcgctagagccagtagcggg tgcagcaata gcagcacccc tcactggtca gcaaaatata 120 attgatccctggattatgaa taattttgtg caggcacctg gtggtgagtt tacagtgtca 180 cccagaaactcccctggtga agtgcttctt aatttagaat taggtccaga aataaatccc 240 tatttggctcaccttgctag aatgtacaat ggttatgcag gcgggtttga agtgcaggta 300 gtcctggctggtaatgcgtt cacagcagga aagataatct ttgca 345 112 345 DNA Norwalk-likeVirus (GII) 112 gctgctccgt ctaacgatgg tgccgccggc ctcgtcccag agatcaacaatgaggcaatg 60 gcgctagacc cagtggcggg tgcagcgata gcagcgcccc tcactggacagcaaaacata 120 attgatccct ggattatgaa taattttgtg caagcacctg gtggtgagtttacagtgtca 180 cctaggaatt cccctggtga agtgcttcta aatttagaat taggcccagaaataaacccc 240 tatctggctc accttgctag gatgtacaat ggttatgcag gtggatttgaagtgcaggta 300 gtcctggctg gaaatgcgtt tacagcagga aaggtgatct ttgca 345 113345 DNA Norwalk-like Virus (GII) 113 gctactccat caaatgatgg tgccgccggcctcgtgccag aaagtaataa tgaggcaatg 60 gctctggaac ccgtggtggg ggcgtctttagccgcccctg tcactggcca aactaatata 120 atagacccct ggattagaac taattttgtccaagccccta atggtgaatt cacagtctcc 180 cctagaaatt cccctggaga gatattggtcaatttggagt tgggtccaga actgaaccct 240 tatctggcac atttagctag gatgtacaatggttatgcgg gtggtatgga ggtgcaagag 300 atgctcgcgg ggaacgcgtt cactgctggcaagatcatct ttgcc 345 114 345 DNA Norwalk-like Virus (GII) 114 gccgctccatctaatgatgg tgcagccggt ctcgtgccag aggtcaacaa cgagacgatg 60 gcactcgaaccggtggctgg ggcttccata gccgcccctc taaccggtca aaacaatgtg 120 atagacccctggattaggat gaactttgtt caagccccaa atggtgaatt tacagtatct 180 ccccgtaattctcctggtga aattctgtta aatttagaat taggtcctga attaaatcca 240 ttcttagcacacctttcaag gatgtacaat ggttatgctg gcggggttga agtgcaggtg 300 ctacttgctgggaacgcgtt cacagcagga aaactagtgt ttgca 345 115 345 DNA Norwalk-likeVirus (GII) 115 gctactccat caaatgatgg tgccgccggc ctcgtgccag aaagtaataatgaggcaatg 60 gctctggaac ccgtggtggg ggcgtcttta gccgcccctg tcactggccaaactaatata 120 atagacccct ggattagaac taattttgtc caagccccta atggtgaattcacagtctcc 180 cctagaaatt cccctgggga gatatcggtc aatttggagt tgggtccagaactggaccct 240 tatctggcac atttagctag gatgtacaat ggttatgcgg gtggtatggaggtgcaagtg 300 atgctcgcgg ggaacgcgtt cactgctggc aaggtcatct ttgcc 345 116345 DNA Norwalk-like Virus (GII) 116 gctactccat caaatgatgg tgccgccggcctcgtgccag aaagtaataa tgaggcaatg 60 gctctggaac ccgtggtggg ggcgtctttagccgcccctg tcactggcca aactaatata 120 atagacccct ggattagaac taattttgtccaagccccca atggtgaatt tacagtttcc 180 cctagaaatt cccctggaga gatattggtcaatttggagt tgggtccaga actgaaccct 240 tatctggcac atttagctag gatgtacaatggttatgcgg gtggtatgga agtgcaagtg 300 atgctcgcgg ggaacgcgtt cactgctggcaagatcatct ttgcc 345 117 345 DNA Norwalk-like Virus (GII) 117 gctgctccgtctaacgatgg tgccgccggc ctcgtcccag agatcaacaa tgaggcaatg 60 gcgctagagccagtggcagg tgcagcaata gcagcacctc tcactggcca gcaaaatata 120 attgatccctggattatgaa taactttgtg caagcacctg gtggtgagtt tacagtgtca 180 cctaggaactcccctggtga agtacttctc aatttagaat taggtccaga aataaaccct 240 tatttggctcaccttgctag gatgtacaat ggttatgcag gtgggtttga ggtgcaggta 300 gtcctggctggaaatgcgtt tacagcagga aaggtgatct ttgca 345 118 343 DNA Norwalk-likeVirus (GII) 118 tgctccatct aatgatggtg ccgccggcct cgtcccagag atcaacaatgaggcaatggc 60 gctagaccca gtggcgggtg cagcgatagc agcacccctc actggtcagcaaaacataat 120 tgatccctgg attatgaata attttgtgca agcacctggt ggtgagtttacagtgtcccc 180 taggaattcc cctggtgaag tgcttcttaa tttggaattg ggcccagaaataaaccctta 240 tttggcccat cttgctagaa tgtataatgg ttatgcaggt ggatttgaagtgcaggtggt 300 cctggctggg aatgcgttca cagcaggaaa gataatcttt gca 343 119343 DNA Norwalk-like Virus (GII) 119 tgctccatct aatgatggtg ccgccggcctcgtcccagag atcaacaatg aggcaatggc 60 gctagaccca gtggcgggtg cagcgatagcagcacccctc actggtcagc aaaacataat 120 tgatccctgg attatgaata attttgtgcaagcacctggt ggtgagttta cagtgtcccc 180 taggaattcc cctggtgaag tgcttcttaatttggaattg ggcccagaaa taaaccctta 240 tttggcccat cttgctagaa tgtataatggttatgcaggt ggatttgaag tgcaggtggt 300 cctggctggg aatgcgttca cagcaggaaagataatcttt gca 343 120 343 DNA Norwalk-like Virus (GII) 120 agctccatcgaatgatggcg cggctggcct cgtaccagag atcaaccatg aggtcatggc 60 catagaacctgttgcagggg cctctttagc agcccctgtc gtaggacaac ttaatataat 120 tgatccctggattagaaata attttgtgca agcccctgct ggagaattta ctgtttcacc 180 tagaaatgctccaggtgaat ttctgttaga tctagagtta ggccctgaat tgaaccccta 240 ccttgctcaccttgcacgca tgtataatgg gcatgcaggt ggtatggagg tgcagatagt 300 gcttgctgggaatgcgttca cagcgggcaa aatcctgttt gca 343 121 32 DNA Artificial Sequenceprobe 121 ccgtcgtggg agggcgatcg caatctcgac gg 32 122 32 DNA ArtificialSequence probe 122 ccgtcgagat tgcgatcgcc ctcccacgac gg 32 123 30 DNAArtificial Sequence probe 123 ccgtcgattg cgatcgccct cccacgacgg 30 124 29DNA Artificial Sequence probe 124 cgtggaattg cgatcgccct ccctccacg 29 12528 DNA Artificial Sequence probe 125 cctgcattgc gatcgccctc ccagcagg 28

1. A method of detecting a virus in a specimen, whereby a Norwalk-likevirus (GII) is detected by using as an index the nucleic acids of acomplimentary nucleic sequence corresponding to the 4851- to5450-positions of the nucleotide sequence of the cDNA of the prototype(standard strain) of the Norwalk-like virus (GII).
 2. A method ofdetecting a virus in a specimen, whereby a Norwalk-like virus (GII) isdetected by using as an index the nucleic acids of complementarynucleotide sequence corresponding to the 4919- to 5389-positions of thenucleotide sequence of the cDNA of the prototype (standard strain) ofthe Norwalk-like virus (GII).
 3. A method of detecting a virus in aspecimen, whereby a Norwalk-like virus (GII) is detected by using as anindex the nucleic acids of a complementary nucleotide sequencecorresponding to the 4988- to 5107-positions of the nucleotide sequenceof the cDNA of the prototype (standard strain) of the Norwalk-like virus(GII).
 4. The method of detecting a virus as described in any of claims1 through 3, wherein the nucleic acids of the complementary nucleotidesequence serving as a detection index and corresponding to a region ofthe nucleotide sequence of the cDNA of the prototype (standard strain)of the Norwalk-like virus (GII) is a gene amplification product obtainedby applying, for attaining gene amplification, gene amplification meansto genes obtained from the specimen, wherein the gene amplificationmeans comprises gene amplification primers engineered on the basis of atleast two nucleotide sequences each having consecutive 10 or more baseswhich are selected from the complementary nucleotide sequence.
 5. Themethod of detecting a virus as described in claim 4, wherein the atleast two complementary nucleotide sequences of consecutive 10 or morebases and serving as a basis for the engineering of gene amplificationprimers have 15 to 30 consecutive bases.
 6. The method of detecting avirus as described in claim 4 or 5, wherein the at least twocomplementary nucleotide sequences of consecutive 10 or more bases andserving as a basis for the engineering of gene amplification primers arenucleotide sequences of consecutive 10 or more bases selected, as abenchmark, from the group consisting of nucleotide sequences of the4988- to 5028-positions of the nucleotide sequence of the cDNA of theprototype (standard strain) of the Norwalk-like virus (GII), the 5080-to 5107-positions thereof, the 4919- to 4941-positions thereof, and the5367- to 5389-positions thereof.
 7. The method of detecting a virus asdescribed in claim 4 or 5, wherein the at least two nucleotide sequencesof consecutive 10 or more bases, the nucleotide sequences serving as abasis for the engineering of gene amplification primers, are the 4988-to 5028-positions and the 5080- to 5107-positions of the nucleotidesequence of the cDNA of the prototype (standard strain) of theNorwalk-like virus (GII).
 8. The method of detecting a virus asdescribed in claim 7, wherein the nucleotide sequences of the geneamplification primers are complementary nucleotide sequencesconstituting the following combinations <1> or <2>: <1> A nucleotidesequence set composed of a nucleotide sequence selected from the groupconsisting of SEQ ID NO: 2, 3 and 4, and one or more sequences selectedfrom among SEQ ID NOs: 7, 8, and 9; <2> A nucleotide sequence setcomposed of a nucleotide sequence of SEQ ID NO: 1, and nucleotidesequence(s) of SEQ ID NO: 5 and/or
 6. 9. The method of detecting a virusas described in any of claims 6 through 8, wherein the geneamplification product to be obtained corresponding to the Norwalk-likevirus (GII) contains at least a complementary nucleotide sequencecorresponding to the 4942- to 5366-positions of the nucleotide sequenceof the cDNA of the prototype of the Norwalk-like virus (GII), and thecomplementary nucleotide sequence is detected as an indication of thepresence of the Norwalk-like virus (GII).
 10. The method of detecting avirus as described in any of claims 6 through 8, wherein the geneamplification product to be obtained corresponding to the Norwalk-likevirus (GII) contains at least a complementary nucleotide sequencecorresponding to the 5042- to 5067-positions of the nucleotide sequenceof the cDNA of the prototype of the Norwalk-like virus (GII), and thecomplementary nucleotide sequence is detected as an indication of thepresence of the Norwalk-like virus (GII).
 11. The method of detecting avirus as described in any of claims 6 through 10, wherein the detectionof gene amplification product with respect to the Norwalk-like virus(GII) is detection of a specific region of the gene amplificationproduct, the detection being performed through at least using a nucleicacid probe for gene detection containing a nucleotide sequencecomplementary to a complementary nucleotide sequence corresponding tothe 5042- to 5067-positions of the nucleotide sequence of the cDNA ofthe prototype (standard strain) of the Norwalk-like virus (GII).
 12. Themethod of detecting a virus as described in any of claims 6 through 11,wherein the nucleic acid probe for gene detection comprises nucleotidesequences of one or more complementary nucleotide sequences selectedfrom the group consisting of SEQ ID NOs: 10, 11, and
 12. 13. The methodof detecting a virus as described in claim 12, wherein the nucleic acidprobe for gene detection is a molecular beacon probe or a Taq-Man probe.14. The method of detecting a virus as described in any of claims 1through 13, which quantitatively detects the Norwalk-like virus (GII).15. A kit for detecting a virus, which is used for performing a methodof detecting a virus as recited in any of claims 1 through 14 andcontaining the following gene amplification primers and/or a nucleicacid probe for gene detection: <1> gene amplification primers,comprising at least two nucleotide sequences of consecutive 10 or morebases falling within the nucleotide sequence of the complementarynucleotide sequence corresponding to the 4919- to 5389-positions of thenucleotide sequence of the cDNA of the prototype (standard strain) ofthe Norwalk-like virus (GII); and <2> a nucleic acid probe for genedetection which is to be used together with the above-described geneamplification primers, which has a nucleotide sequence complementary toa gene amplification product obtained from the gene of the Norwalk-likevirus (GII), and which is labeled with a labeling substance to bedetected.
 16. The kit for detecting a virus as described in claim 15,wherein the nucleotide sequences on the basis of which the geneamplification primers are engineered are complementary nucleotidesequences, one being selected from the 4988- to 5028-positions andanother being selected from the 5080- to 5107-positions of thenucleotide sequence of the cDNA of the prototype (standard strain) ofthe Norwalk-like virus (GII).
 17. The kit for detecting a virus asdescribed in claim 16, wherein the nucleotide sequences of the geneamplification primers are complementary nucleotide sequences whichconstitute either of the following sets <1> or <2>: <1> A nucleotidesequence set composed f a nucleotide sequence selected from the groupconsisting of SEQ ID NO: 2, 3 and 4, and one or more sequences selectedfrom among SEQ ID NOs: 7, 8, and 9; <2> A nucleotide sequence setcomposed of a nucleotide sequence of SEQ ID NO: 1, and nucleotidesequence(s) of SEQ ID NO: 5 and/or
 6. 18. The kit for detecting a virusas described in any of claims 15 to 17, wherein the nucleic acid probefor gene detection comprises a nucleotide sequence complementary to acomplementary nucleotide sequence corresponding to the 5042- to5067-positions of the nucleotide sequence of the cDNA of the prototype(standard strain) of the Norwalk-like virus (GII).
 19. The method ofdetecting a virus as described in any of claims 15 through 18, whereinthe nucleic acid probe for gene detection comprises the nucleotidesequence of one or more complementary nucleotide sequences selected fromthe group consisting of SEQ ID NOs: 10, 11, and
 12. 20. The kit fordetecting a virus as described in any of claims 15 to 19, wherein thenucleic acid probe for gene detection is a molecular beacon probe or aTaq-Man probe.